KR102585071B1 - Guide frame, mounting chamber formed based on guide frame, and method for providing experiment environment of zebrafish using mounting chamber - Google Patents

Abstract

A mounting chamber according to an embodiment of the present disclosure includes a three-dimensional object having a predetermined thickness and including upper and lower surfaces opposing each other, and a main depression formed in the upper surface of the three-dimensional object extending in the longitudinal direction in the first direction. and a first side depression that extends in one direction from the upper surface, has two ends, the first end is connected to the main depression, and is depressed in the upper surface to a depth shallower than the main depression, based on the upper surface. It may be made of a material that has fluidity above a first temperature and loses fluidity below a second temperature.

Description

Guide frame, mounting chamber formed based on the guide frame, method of providing an experimental environment for zebrafish using a mounting chamber {GUIDE FRAME, MOUNTING CHAMBER FORMED BASED ON GUIDE FRAME, AND METHOD FOR PROVIDING EXPERIMENT ENVIRONMENT OF ZEBRAFISH USING MOUNTING CHAMBER}

The present disclosure relates to a mounting chamber formed to place zebrafish embryos at regular intervals, a guide frame used to form the mounting chamber, and a method of providing an experimental environment for zebrafish using the mounting chamber. will be.

Studying the developmental process of vertebrates can be of great help in treating various diseases. In particular, understanding the vascular development process is the basis for the development of treatments that can inhibit angiogenesis in vascular-related diseases and cancer tissues, so active research is being conducted recently. Zebrafish, a tropical freshwater fish, is the simplest vertebrate and is a recently established research model to study vertebrate-specific organ and cell development.

As a research method using such zebrafish, referring to FIG. 1, zebrafish are first classified into male and female (110), the embryos spawned after mating are arranged in a culture dish, and then the gene to be studied is injected. After performing the transformation and nurturing the zebrafish embryo (120), research results can be derived through the process of acquiring and analyzing the image of the embryo (130).

At this time, in order to observe the growing zebrafish embryo, images of each embryo are periodically needed. However, since zebrafish embryos are arranged irregularly in a culture dish (140), it is inevitably difficult and cumbersome to take images of each embryo every time.

The prior art (Patent Publication No. 10-2014-0131754) discloses a structure related to the development of anticancer drugs using zebrafish, but is completely unaware of the inconvenience in the process of acquiring images of zebrafish embryos. Improvement methods for this are also not disclosed.

Therefore, it is necessary to provide an experimental environment in which automated image capture of zebrafish embryos is possible using a mounting chamber that can place zebrafish embryos at regular intervals.

Prior art: Korean Patent Publication No. 10-2014-0131754 (published on November 14, 2014)

One embodiment of the present disclosure provides a mounting chamber capable of arranging zebrafish embryos at regular intervals and a guide frame used to form the mounting chamber, thereby facilitating a mounting chamber capable of arranging zebrafish embryos. The purpose is to enable it to be formed.

One embodiment of the present disclosure provides a mounting chamber capable of arranging zebrafish embryos, thereby automating image taking of each zebrafish embryo by moving the lens of the microscope at predetermined intervals in accordance with the determined arrangement position. The purpose is to provide an environment in which the cultivation process of zebrafish embryos can be easily understood.

An embodiment of the present disclosure provides a mounting chamber including a depression formed on the upper surface of a three-dimensional object and a side depression connected to the depression, and the zebrafish embryo is spaced apart in the depression and then injected through the side depression. By allowing the fixation solution to be delivered to the main depression to cover the zebrafish embryos, the spaced apart zebrafish embryos are fixed, and the zebrafish embryos can be normally cultured in the fixed position based on the culture medium delivered through the fixation solution. The purpose is to make it possible.

One embodiment of the present disclosure may be a method of providing an experimental environment for zebrafish using a mounting chamber formed to place zebrafish embryos at regular intervals, a guide frame used to form the mounting chamber, and a mounting chamber.

An embodiment of the present disclosure includes a plate-shaped plate including upper and lower surfaces opposing each other, a main protrusion formed on the upper surface of the plate extending in a longitudinal direction in a first direction, and a main protrusion extending in a longitudinal direction from the upper surface in one direction. It may be a guide frame that has two ends in an extended form, the first end is connected to the main protrusion, and includes a first side protrusion that protrudes from the upper surface at a lower height than the main protrusion based on the upper surface.

One embodiment of the present disclosure includes a three-dimensional object having a predetermined thickness and including upper and lower surfaces facing each other, a main depression formed in the upper surface of the three-dimensional object extending in the longitudinal direction in a first direction, and a main depression formed on the upper surface of the three-dimensional object, It has two ends extending in one direction, the first end is connected to the main depression, and a first side depression is formed by being depressed in the upper surface to a shallower depth than the main depression, based on the upper surface, The mounting chamber may be made of a material that has fluidity above a first temperature and loses fluidity below a second temperature.

An embodiment of the present disclosure includes a three-dimensional object having a predetermined thickness and including upper and lower surfaces facing each other, a main depression formed in the upper surface of the three-dimensional object extending in the longitudinal direction in a first direction, and the main depression. The portion and the first end are connected and include a first side depression formed by being depressed in the upper surface to a depth shallower than the main depression, with respect to the upper surface, having fluidity above the first temperature and losing fluidity below the second temperature. A mounting chamber made of a material is provided, a zebrafish embryo is injected with a predetermined liquid into the main depression, and the zebrafish embryo is spaced apart, and a mounting chamber formed at the second end of the first side depression is formed. Providing an experimental environment for zebrafish using a mounting chamber, including the step of injecting a fixation solution into the injection port and transferring the injected fixation solution to the main depression to cover the zebrafish embryo by moving along the first side depression. It could be a way.

By providing a guide frame according to an embodiment of the present disclosure, it is possible to easily form a mounting chamber in which zebrafish embryos can be placed at regular intervals.

The mounting chamber according to an embodiment of the present disclosure allows the placement of zebrafish embryos, thereby automatically moving the camera at predetermined intervals to a position determined according to the specifications of the mounting chamber to capture images of each zebrafish embryo. An environment that can be automated can be provided. Through this, embodiments of the present disclosure allow an imaging device (e.g., a microscope) to capture many embryos at an early stage of development (e.g., 1 day, 2 days, 3 days after fertilization, etc.) for zebrafish embryos. Allows images to be acquired quickly and automatically within a set time.

In the method of providing an experimental environment for zebrafish using a mounting chamber according to an embodiment of the present disclosure, zebrafish embryos are spaced apart in depressions in the mounting chamber, and then a fixation solution is injected through a side depression connected to the depression. By allowing the embryos of the zebrafish to be delivered to the main depression to cover them, the embryos of the zebrafish placed at a distance are fixed, and the embryos of the zebrafish can be cultured normally in a fixed position based on the culture medium delivered through the fixation solution. .

Figure 1 is a diagram for explaining a conventional research method using zebrafish.
FIG. 2 is a diagram illustrating a method for researching zebrafish embryos using a guide frame and a mounting chamber according to an embodiment of the present disclosure.
Figure 3 is a perspective view of a guide frame according to an embodiment of the present disclosure.
Figure 4 is a cross-sectional view of a guide frame according to an embodiment of the present disclosure.
5 is a diagram illustrating various embodiments of a guide frame according to an embodiment of the present disclosure.
Figure 6 is a perspective view of a guide frame according to another embodiment of the present disclosure.
7 is a diagram illustrating various embodiments of a guide frame according to another embodiment of the present disclosure.
Figure 8 is a perspective view of a mounting chamber according to an embodiment of the present disclosure.
9 is a cross-sectional view of a mounting chamber according to an embodiment of the present disclosure.
10 is a diagram illustrating various embodiments of a mounting chamber according to an embodiment of the present disclosure.
Figure 11 is a perspective view of a mounting chamber according to another embodiment of the present disclosure.
FIG. 12 is a cross-sectional view of a mounting chamber according to another embodiment of the present disclosure.
13 is a diagram illustrating various embodiments of a mounting chamber according to another embodiment of the present disclosure.
Figure 14 is a flowchart showing a method of providing an experimental environment for zebrafish using a mounting chamber according to an embodiment of the present disclosure.

Hereinafter, embodiments disclosed in the present specification will be described in detail with reference to the attached drawings. However, identical or similar components will be assigned the same reference numbers regardless of reference numerals, and duplicate descriptions thereof will be omitted. The suffixes “module” and “part” for components used in the following description are given or used interchangeably only for the ease of preparing the specification, and do not have distinct meanings or roles in themselves. Additionally, in describing the embodiments disclosed in this specification, if it is determined that detailed descriptions of related known technologies may obscure the gist of the embodiments disclosed in this specification, the detailed descriptions will be omitted. In addition, the attached drawings are only for easy understanding of the embodiments disclosed in this specification, and the technical idea disclosed in this specification is not limited by the attached drawings, and all changes included in the spirit and technical scope of the present invention are not limited. , should be understood to include equivalents or substitutes.

Terms containing ordinal numbers, such as first, second, etc., may be used to describe various components, but the components are not limited by the terms. The above terms are used only for the purpose of distinguishing one component from another.

When a component is said to be "connected" or "connected" to another component, it is understood that it may be directly connected to or connected to the other component, but that other components may exist in between. It should be. On the other hand, when it is mentioned that a component is “directly connected” or “directly connected” to another component, it should be understood that there are no other components in between.

FIG. 2 is a diagram illustrating a method for researching zebrafish embryos using a guide frame and a mounting chamber according to an embodiment of the present disclosure.

Referring to Figure 2, prepare a container (e.g., a culture dish, a schale) containing a guide frame and a set material, and allow the guide frame to be introduced into the material in a sol or gel state. (210). At this time, the upper surface of the guide frame on which protrusions (eg, main protrusions, first and second side protrusions) are formed may be inserted into the material.

Here, the guide frame is a frame used to form a mounting chamber in which zebrafish embryos can be placed at regular intervals, and may be formed, for example, by an injection molding method or a press method. The guide frame may be made of, for example, any of polycarbonate, polypropylene, polyethylene, polystyrene, polyimide, polyethylene terephthalate, polydimethylsiloxane, polylactic-coglycolic acid, polylactic acid, polycaprolactone, glass, acrylic resin and silicone. It may be one, but is not limited to this.

Additionally, the material contained in the container is a material that supports the sol-gel state, and may be, for example, any one of agarose, hyaluronic acid, poloxamer, alginate, gelatin, collagen, and fibrin gel, but is not limited thereto.

The material contained in the container changes to a gel state, creating a three-dimensional object with depressions (e.g., main depression, first and second side depressions) formed at the top according to the shape of the protrusion formed on the upper surface of the guide frame, that is, a mounting chamber. It can be (220).

In one embodiment, when the material in the container is in a sol state, the guide frame is retracted, and the guide frame changes to a gel state in the retracted state, thereby forming a mounting chamber in the form of a solidified gel with a depression formed. At this time, the fixing part 301 may be at a height that allows it to land on the bottom of the container and form a depression in the sol-state material while supporting the guide frame. Alternatively, with a guide frame placed at the bottom of the container, a sol-state material is introduced into the container (the sol-state material is introduced above the height from the bottom of the container to the top of the guide frame to form a depression), and the sol-state material is introduced into the container. The material changes into a gel state, so that a mounting chamber in the form of a solidified gel with a depression formed can be formed.

In another embodiment, when the material in the container is changed to a gel, the guide frame may be inserted to form a gel-shaped mounting chamber with a depression formed. The zebrafish embryo 230 contained in another container is transferred to the pipette 240. Through, it can be placed in the mounting chamber (250). At this time, the zebrafish embryos can be arranged in the mounting chamber.

The mounting chamber can be mounted on the microscope (260). At this time, the mounting chamber without a container may be mounted on the microscope, but the present invention is not limited to this, and the mounting chamber may be mounted on the microscope while contained in a container.

As the zebrafish embryos are arrayed and placed in the mounting chamber, the microscope can easily image each zebrafish embryo by moving the lens (camera, or stage on which the mounting chamber is mounted) of the microscope according to the position of the array. You can shoot. Additionally, by automating lens movement and image taking, the microscope allows for zebrafish embryos to be acquired periodically during each early developmental stage (e.g., day 1, day 2, day 3 after fertilization, etc.). You can conveniently capture multiple images.

Figure 3 is a perspective view of a guide frame according to an embodiment of the present disclosure.

Referring to FIG. 3 , the guide frame 300 may include a plate-shaped plate 310, a main protrusion 320, and a first side protrusion 330.

The plate-shaped plate 310 may include an upper surface 311 and a lower surface facing each other.

The main protrusion 320 may be formed to protrude from the upper surface 311 of the plate 310 and extend in the longitudinal direction in the first direction 313. Here, the upper surface of the main protrusion 320 in the protruding direction may be a flat surface 321 or may be formed as a curved surface 322 curved in a direction opposite to the upper surface 311.

The first side protrusion 330 extends from the upper surface 311 in one direction and may have two ends 331 and 332. The first side protrusion 330 has a first end 331 connected to the main protrusion 320, and may be formed to protrude from the upper surface 311 at a lower height than the main protrusion 320 with respect to the upper surface 311. there is. This means that, within the mounting chamber formed through the guide frame 300, the main depression formed corresponding to the main protrusion 320 is formed at a deeper depth than the first side depression formed corresponding to the first side protrusion 330. By doing so, the fixing solution injected into the first side depression can be well delivered to the main depression. Here, the fixation solution is injected into the main depression of the zebrafish embryo with a predetermined liquid, for example, through a pipette, so that the embryo is covered after being placed in the main depression, thereby fixing the embryo without moving. Allows it to be located at the designated point. The fixative solution is a material that can change from a sol to a gel by cooling, and has porosity in the gel state, allowing the culture medium to supply nutrients to zebrafish. This fixation solution may be, for example, any one of agarose, hyaluronic acid, poloxamer, alginate, gelatin, collagen, and fibrin gel, but is not limited thereto.

Here, one or more first side protrusions 330 may be connected to one main protrusion 320.

At this time, the cross section (section A-A') of the guide frame 300 including the main protrusion 320 in (a) of FIG. 4 is as shown in (b) of FIG. 4, and ( The cross section (section B-B') of the guide frame 300 including the main protrusion 320 and the first side protrusion 330 in a) is as shown in (c) of FIG. 4.

In another embodiment, the guide frame 300 is attached to each of the second ends 332 of the plurality of first side protrusions 330, as shown in (a), (b), and (c) of FIG. 5. It may further include a connected second side protrusion 340. This means that within the mounting chamber formed through the guide frame 300, the fixing solution injected through the second side depression formed corresponding to the second side protrusion 340 is delivered to the plurality of first side depressions and finally to the main side depression. Allows it to be delivered to the depression.

As an example, the second side protrusion 340 has a shape extending in the first longitudinal direction and is formed parallel to the main protrusion 320, and the second ends 332 of the first side protrusion 330 Each can be connected vertically.

Specifically, the second side protrusion 340 may be formed to protrude from the upper surface 311 at the same height as the first side protrusion 330 based on the upper surface 311, but is not limited to this and the first side protrusion ( 330) can be formed at a height higher than that.

Additionally, the guide frame 300 may include first side protrusions 330 of various shapes. For example, the guide frame 300 may include a first side protrusion 330-1 on which an injection protrusion 350 is formed, as shown in (a) of FIG. 5. Here, the injection protrusion 350 may have a circular protruding shape, but is not limited thereto. The formation of the injection protrusion 350 allows the fixing solution to be injected into the injection port formed corresponding to the injection protrusion 350 in the mounting chamber formed through the guide frame 300, so that the fixation solution injected into the injection port is injected into the injection port. By allowing the fixing solution to flow into the relatively narrow first side depression, it is possible to inject the fixing solution more easily than when a separate injection port is not formed, and the fixing solution can easily flow into the first side recess.

As shown in (b) of FIG. 5, the guide frame 300 may include a first side protrusion 330-2 formed with an inclined upper surface. At this time, the upper surface of the first side protrusion 330-2 in the protrusion direction may be formed to protrude from the upper surface 311 so that the closer it gets to the main protrusion 320, the higher the height becomes relative to the upper surface 311. That is, the upper surface of the first side protrusion 330 - 2 in the protrusion direction may be inclined upward toward the main protrusion 320 . The formation of the first side protrusion 330-2 is achieved by allowing the first side depression formed to correspond to the first side protrusion 330-2 to have an inclination within the mounting chamber formed through the guide frame 300. , allowing the fixing solution injected into the first side depression to easily move to the main depression.

Additionally, the guide frame 300 may include a radial first side protrusion 330-3, as shown in (c) of FIG. 5. At this time, the first ends of the plurality of first side protrusions 330 - 3 may be spaced apart from each other and connected to the main protrusion 320 . Additionally, the plurality of first side protrusions 330 - 3 may have second ends 332 connected to the second side protrusions 340 in a manner that radiates from the second side protrusions 340 . The formation of the first side protrusion 330-3 is such that, within the mounting chamber formed through the guide frame 300, the first side depression formed corresponding to the first side protrusion 330-3 is further oriented in a different direction. By forming a large amount, the fixative solution injected into the first side depression can quickly move to the main depression through multiple paths.

In another embodiment, the guide frame 300 does not extend as one, as shown in (d) of FIG. 5, but has a second side protrusion 340-1 separated to correspond to at least one first side protrusion. It can be formed including. At this time, the upper surface of the second side protrusion 340-1 may have an oval shape, but is not limited thereto.

Additionally, in an embodiment, the guide frame 300 may further include a fixing part 301 and a gripping part 302.

For example, the fixing part 301 may be located at a corner of the upper surface 311 of the plate 310 and may be formed to protrude in the shape of a circular pillar, but is not limited thereto. At this time, the fixing part 301 may protrude from the upper surface 311 at a higher height than the main protruding part 320.

The fixing part 301 may fix the guide frame 300 so that it does not move when the guide frame 300 is inserted into the material contained in the container to form the mounting chamber. Here, the fixing part 301 may be coupled to a fixing part formed in the container, or may be inserted deeper than the main protrusion 320 into the material contained in the container.

For example, the gripper 302 may be formed to protrude in a cylindrical shape from the lower surface of the plate 310, but is not limited thereto.

The gripper 302 may be used when the guide frame 300 is inserted into a material contained in a container or when the guide frame 300 is separated from the mounting chamber as the mounting chamber is formed.

Figure 6 is a perspective view of a guide frame according to another embodiment of the present disclosure.

Referring to FIG. 6 , the guide frame 600 may include a plate-shaped plate 610, a main protrusion 620, a first side protrusion 630, and a second side protrusion 640. The form of each component within the guide frame 600 is the same as that described with reference to FIG. 3, and detailed description thereof will be omitted.

However, the main protrusion 620 includes a plurality of first main protrusions 620-1 and second main protrusions 620-2 formed in parallel and spaced apart from each other at set intervals along a second direction perpendicular to the first direction. It can be included. In addition, there are a plurality of first side protrusions 630, each of which has a first end connected to one side in the longitudinal direction of the first main protrusion 620-1 and a second end connected to the second main protrusion 620-2. ) can be formed to be spaced apart and connected to either side of the longitudinal direction.

Additionally, the guide frame 600 may be formed as shown in FIG. 7 by applying various examples in FIG. 5 .

Figure 8 is a perspective view of a mounting chamber according to an embodiment of the present disclosure.

Referring to FIG. 8 , the mounting chamber 800 may include a three-dimensional object 810, a main depression 820, and a first side depression 830. Here, the mounting chamber 800 may be made of a material that has fluidity above the first temperature and loses fluidity below the second temperature. Having fluidity means maintaining the liquid form, and losing fluidity means solidifying or gelling. The material may include, for example, any one of agarose, hyaluronic acid, poloxamer, alginate, gelatin, collagen, and fibrin gel.

The three-dimensional object 810 may have a predetermined thickness and include an upper surface 811 and a lower surface facing each other. Here, the three-dimensional object 810 may be formed in the same shape as the circumference of a container (for example, a petri dish or a schale), but is not limited thereto, and is shown as a square with a predetermined thickness for convenience of explanation.

The main depression 820 may be formed as a depression in the upper surface 811 of the three-dimensional object 810, extending in the longitudinal direction in the first direction 813. In the main depression 820, zebrafish embryos may be arranged to be spaced apart in the first direction 813, for example. Here, the bottom surface of the main depression 820 in the depression direction may be a flat surface 821 or may be formed as a curved surface 822 curved in the opposite direction to the upper surface 811.

The first side depression 830 extends from the upper surface 811 in one direction and may have two end portions 831 and 832. Here, the first end 831 of the first side depression 830 may be connected to the main depression 820, and a fixative solution may be injected into the second end 832 of the first side depression 830. An injection port may be formed. The injection port may be the second end 832 itself or include a separate depression that is distinct from the second end 832 (eg, a second side depression, which will be described below).

Specifically, the first side depression 830 has a first end 831 connected to the main depression 820, and, based on the upper surface 811, has a shallower depth than the main depression 820. ), thereby enabling the fixing solution injected into the first side depression 830 to be well delivered to the main depression 820 formed at a deeper depth than the first side depression 830. At this time, one or more first side depressions 830 may be connected to one main depression 820.

The zebrafish embryo placed in the main depression 820 may be located at a point within the main depression 820 connected to the first side depression 830, but is not limited to this and may be located between points. .

At this time, the cross section (section A-A') of the mounting chamber 800 including the main depression 820 in (a) of FIG. 9 is as shown in (b) of FIG. 9, and The cross-section (cut along B-B') of the mounting chamber 800 including the main depression 820 and the first side depression 830 in (a) is as shown in (c) of FIG. 9 . Here, the embryo may be located near the head (e.g., the crown of the head).

In another embodiment, the mounting chamber 800 is connected to each of the second ends 832 of the plurality of first side depressions 830, as shown in (a), (b), and (c) of FIG. 10. By further including a second side depression 840 connected to , the fixing solution injected through the second side depression 840 can be shared by the plurality of first side depressions 830. That is, the fixing solution injected into the second side depression 840 is formed in each of the first side depressions (first side depression_#1, first side depression_#1, first side depression_#1), the ends of which are connected to the second side depression 840. It can be delivered to each point of the main depression 820 connected to (#2, etc.).

As an example, the second side depression 840 has a shape extending in the first longitudinal direction and is formed parallel to the main depression 820, and the second end of the first side depression 830 ( 832) can be vertically connected to each of them.

Specifically, the second side depression 840 may be formed by being depressed in the upper surface 811 to the same depth as the first side depression 830 with respect to the upper surface 811, but is not limited thereto. It may be formed to have a shallower depth than the side depression 830.

Additionally, the mounting chamber 800 may include a first side depression 830 of various shapes. For example, the mounting chamber 800 may include a first side depression 830-1 in which an injection port 850 is formed, as shown in (a) of FIG. 10. Here, the injection port 850 may be circular (not necessarily in the form of a circle, but may be in the form of a closed curve. In other words, a shape that allows the fixing solution to flow in and not be discharged to the outside is sufficient), but is not limited to this. No. This type of injection port 850 allows the fixing solution to be injected more easily than directly injecting the fixing solution into the injection port 850 using one end of the first side depression 830.

As shown in (b) of FIG. 10, the mounting chamber 800 may include a first side depression 830-2 formed as a bottom surface having an inclination. At this time, the bottom surface in the depression direction of the first side depression 830-2 may be formed by being depressed in the upper surface 811 so that the closer it gets to the main depression 820, the deeper the depth becomes based on the upper surface 811. . That is, the bottom surface of the first side depression 830 - 2 in the depression direction may be formed to slope downward toward the main depression 820 . The first side depression 830-2 has an inclination so that the fixing solution injected into the first side depression 830-2 can easily move to the main depression 820.

Additionally, the mounting chamber 800 may include a radial first side depression 830-3, as shown in (c) of FIG. 10. At this time, the first ends of the plurality of first side depressions 830 - 3 may be spaced apart from each other and connected to the main depression 820 . Additionally, the plurality of first side depressions 830 - 3 may radiate from the second side depression 840 and have second ends connected to the second side depression 840 . A plurality of these first side depressions 830-3 are formed in different directions to quickly move the fixing solution injected into the first side depression 830-3 to the main depression 820 through multiple paths. You can do it.

In another embodiment, the mounting chamber 800 is not extended as one, as shown in (d) of FIG. 10, but the second side depressions 840- are separated corresponding to at least one first side depression. It can be formed including 1).

At this time, the lower surface of the second side depression 840-1 may have an oval shape, but is not limited thereto.

Additionally, the mounting chamber 800 may further include identification information (not shown) of the mounting chamber 800 on one side of the three-dimensional object 810. Here, the identification information of the mounting chamber 800 refers to the type information of the mounting chamber 800 (or the guide frame 600 forming the mounting chamber 800), and is represented by a barcode, QR code, English letters and numbers, etc. It may be displayed as a configured unique number (UID), etc., but is not limited to this. The identification information may be formed as a protrusion on the guide frame 600 and a depression in the mounting chamber 800, or may be printed on the mounting chamber 800 using a separate device.

Using the identification information of the mounting chamber 800, the microscope or camera can confirm the specifications of the corresponding mounting chamber 800. For example, the shape of the repeatedly formed module part (consisting of a main depression and a first side depression (or first and second side depressions)), the number of module parts, and the separation distance between module parts can be confirmed. there is. Here, the shape of the module unit includes, for example, the number of main depressions, the number of first side depressions, the separation distance between the first side depressions, whether or not the second side depressions are included, and the main depression, the first side depression (or, It may include information such as the length and width of the second side depression.

For example, if the identification information of the mounting chamber 800 is 'A-1', five module parts 801 including one main recess and 11 first side recesses are included at intervals of 1 cm. It can mean being formed. Here, the shape of the module portion 801 is specifically, one main depression and 11 first side depressions spaced at 0.5 cm intervals, a main depression with a length of 6 cm and a width of 0.5 cm, a main depression with a length of 2 cm and a width of 0.3 cm. It may include information such as a cm-wide first side depression, but is not limited to this.

Figure 11 is a perspective view of a mounting chamber according to another embodiment of the present disclosure.

Referring to FIG. 11 , the mounting chamber 1100 may include a three-dimensional object 1110, a main depression 1120, a first side depression 1130, and a second side depression 1140.

Since the form of each component in the mounting chamber 1100 is the same as described with reference to FIG. 7, its details are omitted.

However, the main depression 1120 includes a first main depression 1120-1 and a second main depression 1120-1, which are formed in plurality and spaced apart from each other in parallel at a set interval along the second direction perpendicular to the first direction. 2) may be included. In addition, there are a plurality of first side depressions 1130, each of which has a first end connected to one side in the longitudinal direction of the first main depression 1120-1, and a second end connected to the second main depression ( 1120-2) can be formed to be spaced apart and connected to either side of the longitudinal direction.

At this time, the cross section (section A-A') of the mounting chamber 1100 including the main depression 1120 in (a) of FIG. 12 is as shown in (b) of FIG. 12, and The cross-section (cut along B-B') of the mounting chamber 1100 including the main depression 1120 and the first side depression 1130 in (a) is as shown in (c) of FIG. .

Additionally, the mounting chamber 1100 may be formed as shown in FIG. 13 by applying various examples in FIG. 10 .

Figure 14 is a flowchart showing a method of providing an experimental environment for zebrafish using a mounting chamber according to an embodiment of the present disclosure.

Referring to Figure 14, at step 1410, a mounting chamber may be provided. The mounting chamber may be formed including a three-dimensional body, a main depression, and a first side depression. A three-dimensional object may have a predetermined thickness and include upper and lower surfaces that face each other. The main depression may be formed in a depression shape on the upper surface of the three-dimensional object extending in the longitudinal direction, which is the first direction. The first side depression may be formed by having a first end connected to the main depression and being depressed in the upper surface to a shallower depth than the main depression, based on the upper surface.

This mounting chamber may be made of a material that has fluidity above the first temperature and loses fluidity below the second temperature. Having fluidity means maintaining the liquid form, and losing fluidity means solidifying or gelling. The material is a material that can change from a sol to a gel by cooling, and may include, for example, any one of agarose, hyaluronic acid, poloxamer, alginate, gelatin, collagen, and fibrin gel.

In step 1420, the zebrafish embryo is injected along with a predetermined liquid into the main depression in the mounting chamber, so that the zebrafish embryo can be spaced apart. At this time, the zebrafish embryo can be placed in the direction of extension of the main depression. In an embodiment, the main depression may include a plurality of first and second main depressions formed in parallel and spaced apart from each other at a set interval along a second direction perpendicular to the first direction. At this time, the first side depressions are plural, and each first end is connected to one side in the longitudinal direction of the first main depression, and the second end is connected to one side in the longitudinal direction of the second main depression. It can be formed spaced apart.

In step 1430, the fixing solution may be injected into an injection hole formed at the second end of the first side depression in the mounting chamber.

In step 1440, the fixation solution injected into the injection port in the mounting chamber may move along the first side depression and be delivered to the main depression to cover the zebrafish embryo.

Here, when the mounting chamber is connected to each of the second ends of the plurality of first side depressions and further includes a second side depression formed by recessing the upper surface to the same depth as the first side depression as an injection port. , the fixative solution injected into the injection port (second side depression) may move along the first side depression and be delivered to the main depression. At this time, the second side depression has a shape extending in the first longitudinal direction and is formed parallel to the main depression, and may be vertically connected to each of the second ends of the first side depression.

In the specification (particularly in the claims) of the present disclosure, the use of the term “above” and similar referential terms may refer to both the singular and the plural. In addition, when a range is described in the present disclosure, the invention includes the application of individual values within the range (unless there is a statement to the contrary), and each individual value constituting the range is described in the detailed description of the invention. It's the same.

Unless there is an explicit order or description to the contrary regarding the steps constituting the method according to the present disclosure, the steps may be performed in any suitable order. The present disclosure is not necessarily limited by the order of description of the steps above. The use of any examples or illustrative terms (e.g., etc.) in the present disclosure is merely to describe the present disclosure in detail, and unless limited by the claims, the scope of the present disclosure is limited by the examples or illustrative terms. It doesn't work. In addition, those skilled in the art will recognize that various modifications, combinations and changes may be made according to design conditions and factors within the scope of the appended claims or their equivalents.

Therefore, the spirit of the present disclosure should not be limited to the above-described embodiments, and the scope of the patent claims described below as well as all scopes equivalent to or equivalently changed from the claims are within the scope of the spirit of the present disclosure. It will be said to belong to

300: Guide frame
310: plate
320: main protrusion
330: first side protrusion
340: second side protrusion

Claims (18)

A plate-shaped plate including upper and lower surfaces opposing each other;
a main protrusion formed on the upper surface of the plate and extending in a longitudinal direction in a first direction; and
A first side protrusion has two ends extending in one direction from the upper surface, the first end is connected to the main protrusion, and protrudes from the upper surface at a lower height than the main protrusion with respect to the upper surface. Including,
The main protrusion includes a plurality of first main protrusions and second main protrusions formed in parallel and spaced apart from each other at a set interval along a second direction perpendicular to the first direction,
The first side protrusions are plural, each of which has a first end connected to one side in the longitudinal direction of the first main protrusion, and a second end connected to one side in the longitudinal direction of the second main protrusion. Formed by being spaced apart in a shape,
Guide frame.
delete According to paragraph 1,
The upper surface of the main protrusion in the protrusion direction is flat or is formed as a curved surface curved in the opposite direction to the upper surface,
Guide frame.
According to paragraph 1,
Further comprising a second side protrusion connected to each of the second ends of the plurality of first side protrusions and protruding from the upper surface at the same height as the first side protrusion with respect to the upper surface.
Guide frame.
According to paragraph 4,
The second side protrusion is formed parallel to the main protrusion while extending in the first longitudinal direction, and is vertically connected to each of the second ends of the first side protrusion,
Guide frame.
According to paragraph 4,
The first ends of the plurality of first side protrusions are spaced apart from each other and connected to the main protrusion,
The plurality of first side protrusions radiate from the second side protrusions and the second ends are connected to the second side protrusions,
Guide frame.
According to paragraph 1,
The upper surface of the first side protrusion in the protruding direction is formed to protrude from the upper surface so that the closer it gets to the main protrusion, the higher the height becomes relative to the upper surface.
Guide frame.
a three-dimensional object having a predetermined thickness and including upper and lower surfaces facing each other;
a main depression formed on the upper surface of the three-dimensional object with the depression shape extending in a longitudinal direction in a first direction; and
A second end has two ends extending in one direction from the upper surface, the first end is connected to the main depression, and is formed by being depressed in the upper surface to a depth shallower than the main depression with respect to the upper surface. Includes 1 side depression,
It is composed of a material that has fluidity above a first temperature and loses fluidity below a second temperature,
The main depression includes a plurality of first and second main depressions formed in parallel and spaced apart from each other at a set interval along a second direction perpendicular to the first direction,
The first side depressions are plural, each of which has a first end connected to one side in the longitudinal direction of the first main depression, and a second end connected to one side in the longitudinal direction of the second main depression. Formed by being spaced apart in a shape,
Mounting chamber.
delete According to clause 8,
The bottom surface of the main depression in the depression direction is flat or is formed as a curved surface curved in the opposite direction to the upper surface,
Mounting chamber.
According to clause 8,
It is connected to each of the second ends of the plurality of first side depressions, and further includes a second side depression formed by being depressed in the upper surface to the same depth as the first side depression with respect to the upper surface,
Mounting chamber.
According to clause 11,
The second side depression is formed parallel to the main depression while extending in the first longitudinal direction, and is vertically connected to each of the second ends of the first side depression.
Mounting chamber.
According to clause 11,
The first ends of the plurality of first side depressions are spaced apart from each other and connected to the main depression,
The plurality of first side depressions radiate from the second side depressions and the second ends are connected to the second side depressions,
Mounting chamber.
According to clause 8,
The bottom surface of the first side depression in the depression direction is formed by being depressed in the upper surface so that the closer it gets to the main depression, the deeper the depth becomes based on the upper surface,
Mounting chamber.
According to clause 8,
The mounting chamber is,
The material includes any one of agarose, hyaluronic acid, poloxamer, alginate, gelatin, collagen, and fibrin gel.
Mounting chamber.
According to clause 8,
Further comprising identification information of the mounting chamber on one side of the three-dimensional object,
Mounting chamber.
A three-dimensional object having a predetermined thickness and including an upper and lower surface facing each other, a main depression formed in the upper surface of the three-dimensional object and the depression shape extending in the longitudinal direction in a first direction, the main depression and a first end is connected, and includes a first side depression formed by recessing the upper surface to a shallower depth than the main depression, based on the upper surface, and is made of a material that has fluidity above a first temperature and loses fluidity below a second temperature. providing a configured mounting chamber;
Injecting zebrafish embryos along with a predetermined liquid into the main depression, so that the zebrafish embryos are spaced apart;
injecting a fixing solution into an injection hole formed at a second end of the first side depression; and
Comprising the step of delivering the injected fixative solution to the main depression so that it moves along the first side depression and covers the zebrafish embryo.
Method for providing an experimental environment for zebrafish using a mounting chamber.
According to clause 17,
The first side depressions are plural,
The injection port is connected to each of the second ends of the plurality of first side depressions, and the mounting chamber is a second side depression formed by being depressed in the upper surface to the same depth as the first side depression with respect to the upper surface. Contains more wealth,
The step of delivering to the main depression is,
Comprising the step of moving the fixing solution injected into the second side depression along the first side depression and transferring it to the main depression,
Method for providing an experimental environment for zebrafish using a mounting chamber.

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