CN215627966U - Mould, gel support body and culture dish device - Google Patents

Abstract

The utility model provides a mould, a gel support body and a culture dish device, wherein the gel support body is used for fish egg embryo microinjection, and the mould comprises a template body and a plurality of convex blocks which are convexly arranged along the surface of the template body in an extending way, and the convex blocks are arranged at intervals; the lug is used for forming an elastic groove on the gel support body, and the groove is used for placing a roe embryo; wherein: the roe embryo leans on the inner wall terminal surface in the slot along width direction and the slot block that corresponds, along length direction's one end, and the space of inserting the needle of being favorable to the syringe needle is reserved to the other end. The gel support body is manufactured by a mould. The scheme can realize the complete positioning and supporting of the embryo of the medaka egg on the arch back. During microinjection, the injection needle can penetrate the membrane wall of the embryo of the medaka egg on the back of the bow, so that slipping of the needle head during needle insertion is avoided, and the embryo of the medaka egg on the back of the bow can not be deformed. When the injection needle is pulled out, the embryo can not be taken out, and the efficiency and the precision of microinjection are improved.

Description

Mould, gel support body and culture dish device

Technical Field

The utility model relates to the technical field of medical instruments, in particular to a mold, a gel support body and a culture dish device.

Background

The existing microinjection bracket is widely applied to microinjection of egg membranes of biological zebrafish or Japanese medaka. The support is only suitable for microinjection of a soft roe culture dish with the roe diameter of about 1 mm. When the arch-backed medaka roe which is harder than the zebrafish or Japanese medaka roe membrane is injected, the roe embryo is only supported by two opposite forces in the injection direction and the support force of the support, and the roe embryo cannot be completely fixed. The phenomena that eggs of medaka bow slip during microinjection, an injection needle is difficult to insert into embryos of the eggs to complete injection, or the embryos of the eggs are often taken out together and can only be manually separated when the injection needle is withdrawn are caused. The risk of damaging the roe embryo and the injection needle can be brought in the experimental process, and the injection efficiency is greatly influenced.

Chinese patent application CN206375898U describes a fixing device for rapid roe microinjection, which comprises a culture dish template, wherein a plurality of hemispherical protrusions or a plurality of bar-shaped protrusions arranged in parallel for fixing roe are arranged on the lower surface of the template. The fixing device only aims at soft common roe embryos with small roes, and the membranes of the eggs of similar embryos of the medaka with an arch back are very hard, so that the fixing device cannot be positioned and microinjected.

SUMMERY OF THE UTILITY MODEL

In view of the above, the present invention provides a mold, a gel support body and a culture dish device to overcome the disadvantages of the prior art.

The utility model provides the following technical scheme:

in a first aspect, the mold is used for manufacturing a fish egg embryo microinjection gel support body, and comprises a mold body and a plurality of convex blocks, wherein the convex blocks are arranged on the surface of the mold body in a protruding mode, the convex blocks are arranged at intervals and used for forming elastic grooves in the gel support body, the grooves are used for containing fish egg embryos, one convex block corresponds to the position of one fish egg embryo, the width of each convex block is not larger than the diameter of the corresponding fish egg embryo, and the length of each convex block is larger than the diameter of the corresponding fish egg embryo.

In this embodiment, the value of the ratio of the length of the bump to the width of the bump is threshold δ e [7, 12 ].

In this embodiment, the width of the bump is 0.7mm, and the length of the bump ranges from 5mm to 8 mm.

In this embodiment, the bumps are arranged at intervals in groups along the width direction and/or the length direction.

In this embodiment, the bumps are uniformly spaced, and the bumps are arranged in a rectangular array.

In this embodiment, the surface of the template body is adapted to the arrangement shape of the plurality of bumps, the template body is used for forming a water storage tank on the gel support body, and the water storage tank is used for providing a liquid environment for embryos during microinjection.

In this embodiment, the side of the template body extends outwards to form a handle for demolding the mold.

In a second aspect, the utility model further provides a gel support body for microinjection of roe embryos, wherein the gel support body is manufactured by the mold, and comprises a gel body formed corresponding to the template body and a groove formed corresponding to the bump.

In this embodiment, the gel support body is made of an elastomer material.

In a third aspect, the utility model further provides a culture dish device, which comprises a culture dish, gel, a measuring cup for containing the gel and the mold, wherein the mold is placed in the culture dish and is molded into a gel support body through casting of the gel; the gel support body comprises a gel body corresponding to the template body and a groove corresponding to the bump.

The embodiment of the utility model has the following advantages:

(1) a mould is used for manufacturing a fish egg embryo microinjection gel bracket body and comprises a template body and a plurality of lugs which are arranged at intervals and used for forming elastic grooves on the gel bracket body and placing fish egg embryos. The fish egg embryo card is gone into groove one end, and when the syringe needle was injected the fish egg embryo from the groove other end, the thrust effect of syringe needle further leaned on in the inner wall terminal surface of groove along the length direction of groove to the realization is to the complete positioning and the support of fish egg embryo. During microinjection, the injection needle is favorable for penetrating the membrane wall of the roe embryo, so that the injection needle is prevented from slipping during needle insertion, and the roe embryo is prevented from deforming. In a similar way, when the injection needle is pulled away, the embryo can not be taken out, and the efficiency and the precision of microinjection are improved.

(2) One end of the roe embryo along the length direction of the groove is propped against the end surface of the inner wall of the groove, and the other end is reserved with a needle inserting space which is beneficial to microinjection. According to the size of the needle inserting space, the needle inserting direction and angle of the injection needle are adjusted; the efficiency and the precision of microinjection are improved.

The gel bracket body for fish egg embryo microinjection is integrally formed and manufactured by the die, and has a simple structure. The gel support body comprises a gel body which is correspondingly formed with the template body and an elastic part groove which is correspondingly formed with the convex block, and the groove is used for placing roe embryos.

The culture dish device comprises a culture dish, gel, a graduated measuring cup and a mold, and has similar beneficial effects of the mold.

In order to make the aforementioned objects, features and advantages of the present invention more comprehensible and comprehensible, preferred embodiments accompanied with figures are described in detail below.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 shows a schematic view of a mold structure according to an embodiment of the present invention;

FIG. 2 shows a schematic side view of FIG. 1;

FIG. 3 is a graph showing the relationship between the direction and angle of insertion of an injection needle and the depth and length of a groove and the diameter of an embryo of an arch-backed medaka egg during microinjection according to an embodiment of the present invention;

FIG. 4 is a schematic structural diagram of a bump according to another embodiment of the present invention;

FIG. 5 is a schematic structural diagram illustrating a structure of a groove partition corresponding to each bump according to another embodiment of the present invention;

FIG. 6 shows a process diagram of a culture dish arrangement according to an embodiment of the utility model.

The reference numbers illustrate:

1-a template body; 2-a bump; 3-a groove; 4-roe embryos; 5-injection needle; 6-a handle; 7-molding; 8-gel; 9-culture dish; 10-measuring cup; 11-a gel scaffold body; 12-gel bulk.

Detailed Description

Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.

It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. In contrast, when an element is referred to as being "directly on" another element, there are no intervening elements present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only.

In the present invention, unless otherwise expressly stated or limited, the terms "mounted," "connected," "secured," and the like are to be construed broadly and can, for example, be fixedly connected, detachably connected, or integrally formed; can be mechanically or electrically connected; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meanings of the above terms in the present invention can be understood by those skilled in the art according to specific situations.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description of the templates herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the utility model. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Referring to fig. 1 to 3, the utility model provides a mold for manufacturing a fish egg embryo microinjection gel support body 11, which comprises a mold plate body 1 and a plurality of convex blocks 2 which extend and are convexly arranged along the surface of the mold plate body 1, wherein the convex blocks 2 are arranged at intervals; the lug 2 is used for forming an elastic groove 3 on the gel bracket body, and the groove 3 is used for placing a roe embryo; wherein:

roe embryo 4 follows the 3 width direction of slot and the slot 3 block that corresponds, and the one end along 3 length direction of slot leans on in the inner wall terminal surface of slot 3, and the other end reserves the space of inserting the needle that is favorable to syringe needle 5.

In this example, the roe embryo 4 is an arch-backed medaka roe embryo. The channel 3 is made of an elastic gel material. Specifically, the gel solution is cast on the template body 1 and the bumps 2 to form the gel support body 11. The gel bracket body 11 is formed with grooves 3 corresponding to the projections 2 one by one, and the inner surfaces of the grooves 3 are matched with the outer surfaces of the projections 2. The groove 3 is used for placing the embryo of the medaka egg on the back of the bow.

It should be noted that the embryonal body egg membrane hardness of the medaka egg is very high. During microinjection, the embryo of the medaka egg on the back of the arch slips when being positioned on a common bracket body, the injection needle 5 is difficult to insert the embryo to complete injection, or the embryo is taken out together when the injection needle 5 is pulled out of the injection, and only manual separation can be carried out.

In the embodiment, the width of the groove 3 is correspondingly set according to the egg diameter width of the embryo of the oryzias latipes egg, so that the embryo of the oryzias latipes egg can be completely clamped in the corresponding groove 3, and the positioning is reliable. The length of the groove 3 is correspondingly set according to the egg diameter of the embryo of the arch-backed medaka egg and the needle inserting space of the injection needle 5 during microinjection, one end of the embryo of the arch-backed medaka egg along the length direction of the groove 3 is abutted against the end face of the inner wall of the groove 3, and the needle inserting space of the injection needle 5 is reserved at the other end.

It can be understood that, in microinjection, the injection needle 5 is inserted at a position close to the top end of the embryo of the medaka egg on the arch back at the reserved space position of the groove 3. At this time, on one hand, the embryos of the medaka eggs on the arch back are just clamped in the groove 3 along the width direction of the groove 3; on the other hand, the embryo of the medaka egg on the arch back is further abutted against the end face of the inner wall of the groove 3 along the length direction of the groove 3 under the pushing force of the injection needle 5, so that the complete positioning and supporting of the embryo of the medaka egg on the arch back during microinjection are realized. The injection needle 5 can penetrate the membrane wall of the embryo of the medaka egg on the arch back, so that the injection needle 5 is prevented from slipping during the injection and the embryo of the medaka egg on the arch back is prevented from deforming. Similarly, when the injection needle 5 is pulled away, the embryo of the medaka egg cannot be taken out.

Referring to fig. 3, the needle insertion of the injection needle is schematically shown during microinjection. The depth of the self-defined groove 3 is h, and the length of the self-defined groove is b; the egg diameter of the embryo of the medaka egg is eta, and the needle insertion angle of the injection needle 5 is theta.

In another embodiment, the needle insertion position of the injection needle 5 may be set to other positions. Alternatively, the needle is inserted parallel to the angle of the side of the groove 3, or perpendicular to the surface of the groove 3. And will not be described in detail herein.

In a preferred embodiment of the present disclosure, the bump 2 is a solid rectangular block structure. Referring to fig. 4, in another embodiment, the bump 2 may be further provided as a shell-shaped cavity structure. The multilayer bumps 2 can be stacked on each other, so that the storage and the transportation are convenient.

In this embodiment, one roe embryo 4 corresponds to one groove 3, and the width of the bump 2 is not greater than the diameter of the roe corresponding to the roe embryo 4; optionally, the ratio of the length of the bumps 2 to the width of the bumps 2 is thresholded δ e [7, 12 ].

Specifically, one arch-backed medaka ovum embryo is correspondingly placed in one groove 3, and the injected embryo and the non-injected embryo are distinguished. The width of the convex block 2 is slightly less than or equal to the egg diameter of the corresponding fish egg embryo 4, which is beneficial to completely clamping the arch-back medaka egg embryo in the groove 3. And when microinjected, the positioning and supporting force is provided for the embryo of the medaka egg on the arch back.

In this embodiment, the width of the bump 2 is 0.7mm, and the length of the bump 2 is 5mm to 8 mm.

The width of the convex block 2 is 0.7mm which is slightly less than or equal to the egg diameter of the embryo of the medaka egg on the arch back. If the length of the projection 2 is 5mm, the needle inserting space for microinjection is small, and the movable space or the injection direction and the injection angle of the injection needle 5 are selected to be small. On the contrary, if the length of the bump 2 is 8mm, the needle inserting space of the microinjection is larger, the movable space of the injection needle 5 or the injection direction and the injection angle are selected to be larger, and the efficiency and the precision of the microinjection are improved.

In this embodiment, the bumps 2 are arranged at intervals in groups in the width direction and/or the length direction.

In one embodiment of the scheme, the bumps 2 are uniformly arranged at intervals, and the bumps 2 are arranged in a rectangular array, so that the processing is easy. At the beginning of microinjection, the position and the angle of the injection needle 5 are adjusted, the injection needle 5 does not need to be adjusted again in the injection process, and the efficiency and the precision of microinjection are improved.

In another embodiment, a plurality of lugs 2 are arranged at intervals in groups along the width direction, and the grooves 3 corresponding to the lugs 2 can be used for storing roe embryos 4 of different varieties. The groups are arranged at intervals, so that the human eyes can distinguish the groups easily. Alternatively, the grooves 3 may be arranged at intervals in groups in the length direction, or in groups in both the length and width directions.

It will be appreciated that different sizes of the projections 2 (grooves 3) are selected or provided depending on the type of roe embryos 4 to be observed for fully locating corresponding roe embryos 4, for example, arch-backed medaka roe embryos. During microinjection, the embryos of the medaka eggs on the arch back do not slip in the gel scaffold prepared by the mould, the injection needle 5 is easy to insert into the embryos of the medaka eggs on the arch back to complete injection, the embryos of the medaka eggs on the arch back are not taken out together when the injection needle 5 is pulled out of the injection, and manual separation is not needed.

In this embodiment, the width of the protrusion 2 is set correspondingly according to the egg diameter of the embryo of the arch-backed medaka egg, so that the embryo of the arch-backed medaka egg can be completely clamped in the corresponding groove 3, and the positioning is reliable. Furthermore, the arrangement mode of the convex blocks 2 (the grooves 3) can be set according to different groups of the same type of roe embryos 4. In the scheme, the embryos of the medaka eggs on the arch back are only divided into one group, and the grooves 3 are uniformly arranged at intervals. When the embryos of the medaka eggs on the arch back are placed, the embryos are sequentially placed according to the arrangement sequence of the grooves 3, so that the single groove 3 is prevented from being placed in a missing mode or in a multiple mode. In other embodiments, for example, when performing the comparison experiment, the embodiment and the comparative example need to be separately placed, so as to avoid confusion and improve the accuracy and efficiency of the comparison experiment. At this time, the number of groups and the number of experiments according to the examples and comparative examples were set accordingly. For example, in this experiment, the number of repetitions was five. In a single experiment, five groups of custom examples and five groups of comparative examples were used. If the experimental observation time is long, five experiments can be carried out simultaneously; specifically, the grooves 3 corresponding to five groups of examples and five groups of comparative examples in a single experiment are divided into one zone, and the corresponding grooves 3 in each experiment are arranged at intervals in a partitioning manner, so that the arrangement and observation are easy. If the time for experiment observation is short, five experiments can be performed in sequence, and the next experiment is performed after the previous experiment is completed. Thus, the number of the grooves 3 can be reduced, and the size of the entire mold can be reduced.

As a further improvement of the technical scheme, the arrangement mode of the corresponding bumps 2 (grooves 3) can be selected for different varieties of roe embryos 4 according to the embodiment and the comparative example. For example, comparative experiments were observed for zebrafish egg embryos and medaka egg embryos. Referring to fig. 5, the grooves 3 corresponding to the bumps 2 are arranged in a partitioned manner and used for placing zebra fish egg embryos and medaka egg embryos respectively. Specifically, the zebra fish egg embryo is placed in the area I, the medaka egg embryo is placed in the area II, and the area I and the area II are separated by the distance S. The grooves 3 are uniformly arranged among zebra fish egg embryos or medaka egg embryos at intervals, and are easy to place and observe.

In this embodiment, the surface of the template body 1 is adapted to the arrangement shape of the plurality of bumps 2, and the template body 1 is used to form a water storage tank on the gel support body 11, and the water storage tank is used to provide a liquid environment for the embryo during microinjection.

In the scheme, the thickness of the template body 1 is matched with the water storage height required by the liquid environment of the roe embryo 4 during microinjection. The plurality of lugs 2 are arranged in a rectangular shape, and the surface of the template body 1 is of a rectangular structure matched with the rectangular shape. The thickness of the template body 1 is used for storing water, and the template body is adapted according to the water storage height required by the liquid environment of the roe embryo 4 during microinjection. During microinjection, the liquid cover is arranged above the roe embryo 4, so that the roe embryo 4 can be in a liquid environment at any time and can not lose water and die. In the preferred embodiment, the length and width of the template body 1 are both 60mm, and the height is 3 mm.

In this embodiment, the side of the formwork body 1 extends outward to form a handle 6 for demoulding the mould. The handle 6 has the function that the mould can be more conveniently taken out after the gel support is solidified, and the gel support is not easy to damage.

Referring to fig. 1, the present invention further provides a gel support body 11, wherein the gel support body 11 is manufactured by the above-mentioned mold. The gel support body 11 comprises a gel body 12 formed corresponding to the template body 1 and a groove 3 formed corresponding to the bump 2. Referring to fig. 6 and 1, the present invention further provides a culture dish device comprising the above mold 7, gel 8, culture dish 9, and measuring cup 10 for containing gel 8; the mold 7 is placed in a petri dish 9, and is cast and molded into a gel support body 11 through a gel 8. The gel support body 11 includes a gel body 12 corresponding to the template body 1, and grooves 3 corresponding to the projections 2. The groove 3 is used for storing the fish egg embryo 4 for microinjection, and the thickness of the gel body 12 is used for storing water and is adapted according to the water storage height required by the liquid environment of the fish egg embryo 4 during microinjection. During microinjection, the liquid cover is arranged above the roe embryo 4, so that the roe embryo 4 can be in a liquid environment at any time and can not lose water and die.

In the scheme, the gel 8 is an elastic gel prepared from an agarose solution, and the prepared gel body 12 and the grooves 3 are elastic components, so that the roe embryos 4 are easily clamped in the corresponding grooves 3 along the width direction.

The specific manufacturing process of the gel support body 11 is as follows: the measuring cup 10 is filled with 20% agarose solution, poured into the culture dish 9 and then placed in the mold 7. The agarose solution is cooled to form an elastic gel 8, and then a gel support body 11 is cast in the petri dish 9. The corresponding gel body 12 is cast through the template body 1 and the corresponding groove 3 is cast through the bump 2. In this example, the roe embryo 4 is an arch-backed medaka roe embryo with a high membrane hardness. Placing each arch medaka egg embryo in the groove 3 of the corresponding gel bracket body 11. In microinjection, the needle insertion direction and angle of the injection needle 5 are first adjusted. And then introducing the injection solvent into the interior of the medaka ova embryo through the injection needle 5 to prepare for microscopic observation.

In the preferred embodiment, the needle insertion direction and angle of the injection needle 5 are selected according to the length of the groove 3 of the gel support body 11 and the reserved needle insertion space. Alternatively, the injection needle 5 is perpendicular to the surface of the embryo of the medaka egg held on the arch, or the injection needle 5 is inserted parallel to the side of the groove 3. The embryo of the arch-back medaka roe cannot slip in the gel support body 11, the injection needle 5 is easy to insert into the arch-back medaka roe embryo to complete injection, after the injection is completed, the injection needle 5 is pulled out of the embryo during the injection and cannot take the embryo out together, manual separation is not needed, and the precision and the efficiency of microinjection are improved.

In all examples shown and described herein, any particular value should be construed as merely exemplary, and not as a limitation, and thus other examples of example embodiments may have different values.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

The above examples are merely illustrative of several embodiments of the present invention, and the description thereof is more specific and detailed, but not to be construed as limiting the scope of the utility model. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

Claims (10)

1. The mold is characterized by comprising a mold body and a plurality of convex blocks, wherein the convex blocks are convexly arranged on the surface of the mold body in an extending mode, the convex blocks are arranged at intervals and used for forming elastic grooves in the gel support body, the grooves are used for containing roe embryos, one convex block corresponds to the position of one roe embryo, the width of each convex block is not larger than the egg diameter of the corresponding roe embryo, and the length of each convex block is larger than the egg diameter of the corresponding roe embryo.

2. The mold according to claim 1, characterized in that the value threshold δ e [7, 12] of the ratio of the length of the bumps to the width of the bumps.

3. The mold of claim 2, wherein the width of the bump is 0.7mm and the length of the bump is in the range of 5mm to 8 mm.

4. The mold according to claim 1, wherein the bumps are arranged at intervals in groups in the width direction and/or the length direction.

5. The mold of claim 1, wherein the bumps are evenly spaced, and the bumps are arranged in a rectangular array.

6. The mold of claim 1, wherein the surface of the mold plate body is adapted to the arrangement shape of the plurality of projections, and the mold plate body is used to form a reservoir on the gel support body, the reservoir being used to provide a liquid environment for the embryo during microinjection.

7. The mold of any one of claims 1 to 6, wherein the side edges of the template body extend outwardly to provide handles for demolding of the mold.

8. A gel scaffold for microinjection of roe embryos, the gel scaffold being manufactured by the mold according to any one of claims 1 to 7, the gel scaffold comprising a gel body molded in correspondence with the template body, and grooves molded in correspondence with the projections.

9. The gel support of claim 8, wherein the gel support body is made of an elastomer material.

10. A petri dish device, which is characterized by comprising a petri dish, gel, a measuring cup for containing the gel and the mould of any one of claims 1 to 7, wherein the mould is placed in the petri dish, and is cast and molded into a gel support body through the gel; the gel support body comprises a gel body corresponding to the template body and a groove corresponding to the bump.

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