KR20180001507U - Flounder eggs microinjection mold for manufacturing agarose gel - Google Patents

Abstract

The present invention provides a mold for preparing agarose gel for microinjection of flounder embryos capable of efficiently performing microinjection of various sizes of flounder embryos.

Description

{Flounder eggs microinjection mold for manufacturing agarose gel}

The present invention relates to a mold for preparing an agarose gel, and more particularly, to a mold for preparing an agarose gel for microinjection of a flounder embryo.

Zebrafish egg microinjection is a nationally and globally generalized technique, and has about two or three microinjection molds in laboratories that treat zebrafish as an experimental animal model. Zebrafish has a bump for the manufacture of agarose gel molds of different sizes than preparing agarose plates of varying sizes because the egg size varies for each cancer / This is because the use of one agarose plate is efficient in carrying out the experiment. In particular, in the case of microinjection using a flounder embryo, a mold for microinjection in the form of a protrusion for producing a rectangular insertion hole which can be easily positioned because it has a microinjection position is fixed and does not move easily after positioning Should be used. Thus, molds for preparing agarose gels with square protrusions used for microinjection for transgenic fish production have been developed (Martina et al., Nature Protocols 1, 1133 - 1139, 2006).

However, in the case of the above-mentioned prior art, it is inefficient to apply to embryos in which the widths of the projections are uniform and the sizes are slightly different.

The object of the present invention is to provide a mold for preparing agarose gel for microinjection of flounder embryos capable of efficiently performing microinjection of various sizes of flounder embryos. However, such a problem is illustrative, and thus the scope of the present invention is not limited thereto.

According to one aspect of the present invention, a rectangular plate of transparent material; And a plurality of protrusions in the longitudinal direction for producing a hole for fixing the flounder embryo formed on the upper surface of the plate, wherein the plurality of protrusions are arranged in the order from the left to the right, There is provided a mold for preparing an agarose gel for microinjection of a flounder embryo, the width of which is increased or decreased at a constant rate from the right side to the left side.

According to another aspect of the present invention, there is provided a method of preparing a gel, comprising: pouring a melted agarose solution into a mold to form an agarose gel; Placing the mold for preparing agarose gel for microinjection of the flounder embryo as set forth in any one of claims 1 to 6 on the top surface of the agarose gel before the gel is completely hardened; And a step of desorbing the mold for preparing an agarose gel for microinjection of the flounder embryo, after the gelatinous agarose gel is completely hardened, and a method for producing the agarose gel for microinjection of the flounder embryo.

According to the idea of the present invention, the mold for preparing the agarose gel for microinjection of the flounder embryo can be rapidly and efficiently microinjected into various sizes of flounder embryos. Of course, the scope of the present invention is not limited by these effects.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a front view showing a mold for preparing an agarose gel for microinjection of a zebrafish embryo which has been commercialized. FIG.
2 is a photograph showing a mold for preparing an agarose gel for microinjection of a zebrafish embryo which has been commercialized.
3 is a view illustrating a microinjection process using a commercial mold for preparing an agarose gel for microinjection of a zebrafish embryo.
FIG. 4 is a perspective view showing a mold for preparing an agarose gel for microinjection of a zebrafish embryo, which is being commercialized, in which a bump for producing a square-shaped insertion hole is formed in a microinjection mold.
5 is a view showing a mold 100 for preparing an agarose gel for microinjection of a flounder embryo according to certain preferred embodiments of the present invention.
6 is a perspective view showing a mold 100 for preparing an agarose gel for microinjection of a flounder embryo according to certain preferred embodiments of the present invention.
7 is a photograph showing a mold 100 for preparing an agarose gel for microinjection of a flounder embryo with a handle according to another embodiment of the present invention.

Detailed description of the design:

According to one aspect of the present invention, a rectangular plate of transparent material; And a plurality of protrusions in the longitudinal direction for producing a hole for fixing the flounder embryo formed on the upper surface of the plate, wherein the plurality of protrusions are arranged in the order from the left to the right, There is provided a mold for preparing an agarose gel for microinjection of a flounder embryo, the width of which is increased or decreased at a constant rate from the right side to the left side.

In the mold for preparing an agarose gel for microinjection of the flounder embryo, the plurality of protrusions may have the same width with two or three adjacent protrusions as a set, and the width may increase or decrease at a constant rate toward the next set.

In the mold for preparing agarose gel for microinjection of the flounder embryo, the width of the protrusion may be 0.5 to 1.2 mm and the height of the protrusion may be 0.7 to 1.0 mm.

In the mold for producing the agarose gel for microinjection of the flounder embryos, the material of the plate may be glass or plastic, and the resin may be urethane, polyvinyl chloride, polyethylene for example, polyethylene, polystyrene, polypropylene, acrylic, nylon or methacrylic resin (pmma).

In the mold for producing the agarose gel for microinjection of the flounder embryos, the predetermined ratio may be 0.05 to 0.2 mm per protrusion, 0.075 to 0.15 mm or 0.09 to 0.12 mm per protrusion, and in one embodiment of the present invention, 0.1 mm , But the present invention is not limited thereto.

According to another aspect of the present invention, there is provided a method of preparing a gel, comprising: pouring a melted agarose solution into a mold to form an agarose gel; Placing the mold for preparing agarose gel for microinjection of the flounder embryo as set forth in any one of claims 1 to 6 on the top surface of the agarose gel before the gel is completely hardened; And a step of desorbing the mold for preparing an agarose gel for microinjection of the flounder embryo, after the gelatinous agarose gel is completely hardened, and a method for producing the agarose gel for microinjection of the flounder embryo.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

Embodiments of the present invention are provided to more fully illustrate the present invention to those skilled in the art, and the following embodiments may be modified in various ways, The present invention is not limited to the embodiment. Rather, these embodiments are provided so that this disclosure will be more thorough and complete, and will fully convey the concept of the invention to those skilled in the art. In the drawings, the thickness and size of each layer are exaggerated for convenience and clarity of explanation.

It is to be understood that when referring to throughout the document that one component, such as a film, region or substrate, is "on", "connected to", "laminated" or "coupled to" another component It will be appreciated that elements may be directly "on", "connected", "laminated" or "coupled" to another element, or there may be other elements intervening therebetween. On the other hand, when one element is referred to as being "directly on", "directly connected", or "directly coupled" to another element, it is interpreted that there are no other components intervening therebetween do. Like numbers refer to like elements. As used herein, the term "and / or" includes any and all combinations of one or more of the listed items.

In this document, the terms first, second, etc. are used to describe various elements, components, regions, layers and / or sections, but these members, parts, regions, layers and / It is obvious that no. These terms are only used to distinguish one member, component, region, layer or section from another region, layer or section. Thus, a first member, component, region, layer or section described below may refer to a second member, component, region, layer or section without departing from the teachings of the present disclosure.

Also, relative terms such as "top" or "above" and "under" or "below" can be used herein to describe the relationship of certain elements to other elements as illustrated in the Figures. Relative terms are intended to include different orientations of the device in addition to those depicted in the Figures. For example, in the figures the elements are turned over so that the elements depicted as being on the top surface of the other elements are oriented on the bottom surface of the other elements. Thus, the example "top" may include both "under" and "top" directions depending on the particular orientation of the figure. If the elements are oriented in different directions (rotated 90 degrees with respect to the other direction), the relative descriptions used herein can be interpreted accordingly.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms "a," "an," and "the" include singular forms unless the context clearly dictates otherwise. Also, " comprise "and / or" comprising "when used herein should be interpreted as specifying the presence of stated shapes, numbers, steps, operations, elements, elements, and / And does not preclude the presence or addition of one or more other features, integers, operations, elements, elements, and / or groups.

Hereinafter, embodiments of the present invention will be described with reference to the drawings schematically illustrating ideal embodiments of the present invention. In the figures, for example, variations in the shape shown may be expected, depending on manufacturing techniques and / or tolerances. Accordingly, the embodiments of the present invention should not be construed as being limited to any particular shape of the regions shown in this document, but should include, for example, changes in shape resulting from manufacturing.

1 shows a microinjection mold for agarose gel for microinjection of a zebrafish embryo which is commercially available for making grooves in an agarose gel for microinjection and for sorting eggs of a zebrafish. (Zebrafish guide book, https://zfin.org/zf_info/zfbook/chapt5/5.1.html). As shown in the figure, a triangular or triangular shaped bump is formed on the agarose gel to form a triangular or quadrangular groove. Referring to FIG. 2, the structure of the mold for preparing the agarose gel for microinjection of the zebrafish embryo It can be easily understood.

FIG. 3 is a view showing a process of microinjection using a mold for producing agarose gel for microinjection of zebrafish, which is being commercialized. In the case of a mold for preparing microaglyceride agarose gels for microinjection of a zebrafish embryo, if the shape of the furrow is a triangle, the injection needle at the time of microinjection is inside the egg Often occurs when the egg is injected into the cytoplasm of the needle and then the egg is not separated from the needle when the needle is withdrawn. At this time, the injection needle and the injection solution should be prepared again as the injection needle breaks frequently while separating the embryo using a method of pushing with the transparent plastic cover together with the culture medium, and the microinjection volume of the injection needle volume and quantification of the volume of the sample. In order to solve such a problem, as shown in FIG. 4, a bump for manufacturing a rectangular insertion hole is formed in a microfiltration frame, and a zebrafish embryo is inserted into a protrusion for manufacturing the insertion hole A mold was developed, but a protrusion for manufacturing an insertion hole was manufactured without considering the size of the embryo, so that an insertion hole was formed in the agarose gel. After the embedding process, the zebrafish embryo was correctly inserted In other words, the size of fertilized egg differs by 0.7 ~ 1.2 mm for each individual. Therefore, when too large fertilized eggs are inserted in the insertion hole, it is not possible to easily fix the injection site using the weak needle, There is a problem that embryo is not fixed properly.

FIG. 5 shows a configuration of a mold 100 for producing an agarose gel for microinjection of a flounder embryo according to some preferred embodiments of the present invention. As shown in the figure, unlike a mold for microinjection having a uniform size protrusion for manufacturing an agarose gel insertion hole, protrusions for manufacturing an insertion hole having various widths capable of fixing various sizes of embryo according to the size of embryo (105) are formed. Thus, it is possible to perform experiments efficiently. In the case of the flounder embryo, since the needle does not easily fall out after injection due to the nature of the cholion, it is necessary to use an agarose plate having protrusions (105) for producing a rectangular insertion hole, In the case of conventional products having protrusions 105 for manufacturing the same size of insertion holes on petridish, it is necessary to manufacture molds for each size and to manufacture insertion holes corresponding to the size of the scattered embryos. An agarose plate having protrusions 105 should be used. However, in the present invention, the mold 100 for preparing an egg-shaped agarose gel for microinjection of a flounder embryo is provided with protrusions 105 for manufacturing a rectangular insertion hole for fixing the size of the embryo to a size that can be used for petridish (ID100 mm) And the width of the protrusion 105 for manufacturing an insertion hole can be adjusted to 0.5 to 1.0 mm so as to be applicable to flounder embryos of various sizes and can be manufactured to have a height of 0.7 to 1.0 mm. 6, it is easy to understand the structure of the mold for preparing the agarose gel for microinjection of the flounder embryo of the present invention, in which the protrusions 105 for producing insertion holes having various widths are formed. In the present invention, for the sake of easy understanding of the structure of the mold for preparing the agarose gel for microinjection of the flounder embryos, the configuration of the protrusions 105 for the production of six insertion holes is shown for convenience, and the width is 0.7 to 0.9 and the height is 0.9. And can be appropriately adjusted according to the purpose and work environment of the researcher. In addition, the length of the protrusion 105 for manufacturing the insertion hole is also 55 mm, which can be appropriately adjusted according to the optimum configuration (see FIG. 5) for experiment execution and also according to the researcher's purpose and work environment.

In addition, as shown in FIG. 7, for the sake of research efficiency, one surface of the mold for microinjection is made of protrusions 105 for the production of a rectangular insertion hole, and the ease of handling in the manufacture of agarose gel A knob or the like may be constructed for the purpose.

Using the mold (100) for preparing the flounder embryo for microinjection of flounder embryos, it is possible to easily fix the microinjection position according to the size of the embryo during microinjection using various sizes of flounder embryos, so that rapid and efficient microinjection It is possible.

Although the present invention has been described with reference to the embodiments shown in the drawings, it is to be understood that various modifications and equivalents may be made thereto by those skilled in the art. Therefore, the true scope of technical protection of the present invention should be determined by the technical idea of the appended utility model registration claim scope.

100: mold for producing agarose gel for microinjection of flounder embryos
105: projection for manufacturing an insertion hole

Claims (8)

Square plate of transparent material; And
And two or more protrusions in the length direction for producing a hole for fixing a flounder embryo formed on an upper surface of the plate, wherein the mold for preparing an egg-
Wherein the plurality of protrusions are increased or decreased in width from a left side to a right side or from a right side to a left side at a constant rate. The method according to claim 1,
Wherein the plurality of protrusions have the same width with two or three adjacent protrusions as a set, and the width is increased or decreased at a constant rate toward the next set, the mold for preparing the olive floss gel for microinjection. The method according to claim 1,
Wherein the width of the protrusions is 0.5 to 1.2 mm, wherein the mold for preparing the agarose gel for microinjection of the flounder embryo. The method according to claim 1,
The height of the protrusions is 0.7 to 1.0 mm. The mold for preparing the agarose gel for microinjection of the flounder embryo. The method according to claim 1,
Wherein the material of the plate is glass or plastic. The mold for producing an agarose gel for microinjection of a flounder embryo. 6. The method of claim 5,
The resin may be selected from the group consisting of urethane, polyvinyl chloride, polyethylene, polystyrene, polypropylene, acrylic, nylon or methacrylic resin (pmma) , Mold for preparing agarose gel for microinjection of flounder embryo. 7. The method according to any one of claims 1 to 6,
Wherein the constant rate is 0.05 to 0.2 mm per lobe, wherein the mold for preparing the agarose gel for microinjection of the flounder embryo. Pouring the molten agarose solution into a mold to form an agarose gel; And
Placing the mold for preparing agarose gel for microinjection of the flounder embryo as set forth in any one of claims 1 to 6 on the top surface of the agarose gel before the gel is completely hardened; And
The method for preparing agarose gel for microinjection of olive flounder embryo, comprising the step of desalting the mold for preparing the agarose gel for microinjection of the flounder embryo after the gelatin agarose gel is completely hardened.

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