阅读说明：本技术 小鼠双染透明骨骼塑封包埋标本制作方法 (Method for manufacturing mouse double-dyeing transparent skeleton plastic package embedding specimen ) 是由 李雁羽 王磊 刘天润 吴启 徐莉 于 2021-08-09 设计创作，主要内容包括：本发明公开了小鼠双染透明骨骼塑封包埋标本制作方法,依次包括材料准备、梯度乙醇脱水固定、丙酮脱脂、混合染色剂染色、复水、预透明、脱色、梯度甘油透明处理、塑化、UV胶包埋,最终获得小鼠双染透明骨骼塑化染色包埋标本。本发明同传统的透明骨骼标本相比,这种标本采取先塑化后包埋的解决方案,有效地解决了传统透明骨骼标本必须保存在灌注甘油的容器之中,不易于携带和展示且会发生脱色的问题。采用本方法制作出的小鼠透明骨骼塑化包埋标本兼具透明骨骼标本,塑化标本,包埋标本三者之长,便于展示,携带,保存,运输。(The invention discloses a method for manufacturing a mouse double-dyed transparent skeleton plastic package embedded specimen, which sequentially comprises the steps of material preparation, gradient ethanol dehydration and fixation, acetone degreasing, mixed coloring agent dyeing, rehydration, pre-transparence, decoloration, gradient glycerol transparency treatment, plasticization and UV (ultraviolet) glue embedding, and finally the mouse double-dyed transparent skeleton plastic package embedded specimen is obtained. Compared with the traditional transparent bone specimen, the specimen adopts the solution of plasticizing and embedding, and effectively solves the problems that the traditional transparent bone specimen is required to be stored in a container filled with glycerol, is not easy to carry and display and can be decolorized. The mouse transparent skeleton plasticized and embedded specimen manufactured by the method has the advantages of being transparent, plasticized and embedded, and is convenient to display, carry, store and transport.)

1. The preparation method of the mouse double-dyed transparent skeleton plastic package embedded specimen is characterized by sequentially comprising the steps of material preparation, gradient ethanol dehydration and fixation, acetone degreasing, mixed staining agent dyeing, rehydration, pre-transparence, decoloration, gradient glycerol transparence treatment, plasticization and UV glue embedding, and finally obtaining the mouse double-dyed transparent skeleton plastic package embedded specimen.

2. The method for preparing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen according to claim 1, wherein the acetone gradient ethanol dehydration fixation step comprises: and (3) soaking the mice subjected to the material preparation step in ethanol with the concentration gradient from low to high in sequence for dehydration and fixation treatment.

3. The method for preparing a mouse double-staining transparent skeleton plastic-packaged embedded specimen according to claim 1, wherein the acetone degreasing step comprises: taking out the mouse subjected to gradient ethanol dehydration fixation treatment from ethanol, soaking in anhydrous acetone, performing degreasing treatment, taking out the mouse from the anhydrous acetone after 20-28h, and soaking in water for 10-14 h.

4. The method for preparing a mouse double-dyed transparent bone plastic-sealed embedded specimen according to claim 1, wherein the dyeing step of the mixed dyeing agent comprises the following steps: soaking a mouse degreased by acetone in a mixed dye solution for 20-28h, wherein the mixed dye solution is a mixture formed by sequentially mixing alizarin red stock solution, aliskiren blue stock solution, acetic acid and 75% ethanol solution;

preferably, the volume ratio of the alizarin red stock solution to the alisnew blue stock solution to the acetic acid to the 75% ethanol solution is 1:0.8-1.2:0.8-1.2: 16-18;

preferably, the alizarin red stock solution is a mixture formed by mixing 25mg of alizarin red and 25ml of 95% ethanol solution by volume, and the alisnew blue stock solution is a mixture formed by mixing 75mg of alisnew blue and 25ml of 95% ethanol solution by volume.

5. The method for preparing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen according to claim 4, wherein the rehydration step is as follows: and taking the mice subjected to the dyeing step of the mixed dyeing agent out of the mixed dyeing agent, and sequentially soaking the mice in absolute ethyl alcohol and pure water for 10-14 hours respectively.

6. The method for preparing a mouse double-dyeing transparent bone plastic-package embedded specimen according to claim 1, wherein the pre-transparentizing step comprises: soaking the mice subjected to the rehydration step in a potassium hydroxide solution with the mass fraction of 2%, wherein the standard for stopping soaking is that the complete spines of the mice can be seen through semitransparent mouse muscles, the hard bones of the mice are light pink, and the cartilages of the mice are blue.

7. The method for preparing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen according to claim 1, wherein the decoloring step comprises the following steps: soaking the mice subjected to the pre-transparency step in a destaining solution I for 1-3d, and then soaking the mice in a destaining solution II for 1-3 d;

preferably, the transparent liquid I is a mixture of a potassium hydroxide solution with the mass fraction of 1% and pure glycerol which are uniformly mixed in a volume ratio of 1:0.8-1.2, and the transparent liquid II is a mixture of water and pure glycerol which are uniformly mixed in a volume ratio of 1: 0.8-1.2.

8. The method for preparing a mouse double-dyeing transparent bone plastic-package embedded specimen according to claim 1, wherein the gradient glycerol transparent treatment comprises the following steps: and (3) sequentially immersing the mice subjected to the decolorization step into glycerol with the volume fraction increasing gradually for 1-3 days respectively, and then immersing the mice into pure glycerol until the muscles are colorless and transparent and the bone structures are clearly visible.

9. The method for preparing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen according to claim 1, wherein the plasticizing step comprises: selecting PEG-600 as a plasticizer, adding thymol crystals into a potassium hydroxide solution with the mass fraction of 0.5%, fully mixing, mixing the potassium hydroxide solution added with the thymol crystals and the PEG-600 according to a certain proportion to respectively prepare a mixture with the volume concentration of 15%, 25%, 50%, 75% and 100%, sequentially putting the mice into the mixture for soaking for 1-2d when the concentration is 15%, and then soaking for 2-3d when each gradient is followed, so that the glycerol in the PEG-600 can be completely replaced by the glycerol to finish plasticization.

10. The method for preparing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen according to claim 1, wherein the UV glue embedding step comprises: pouring UV glue into a silica gel mold, stopping pouring when the UV glue does not cover 1cm of the bottom of the mold, placing the mold in an ultraviolet irradiation environment, placing a mouse subjected to a plasticizing step into the mold after the UV glue on the bottom layer is solidified, adjusting the posture, then pouring the UV glue until the mouse does not cover about 1cm of a specimen, placing the mold in the ultraviolet irradiation environment, demolding and taking out the embedded specimen after the UV glue is completely solidified, and polishing and finishing to obtain the mouse double-dyed transparent skeleton plastic package embedded specimen.

Technical Field

The invention relates to the technical field of specimen preparation, in particular to a preparation method of a mouse double-dyed transparent skeleton plastic package embedded specimen.

Background

Animal bone specimen is a part of natural science popularization teaching, and mouse bone specimen is an important teaching aid for demonstrating the structure of mammal bone specimen. The traditional mouse bone system display samples comprise two types, one type is a bone sample with all the muscle of the mouse removed, the other type is a bone sample with the muscle of the mouse subjected to transparentization treatment and a specific staining agent, and then the muscle of the sample is subjected to decoloration treatment, so that the bone morphology and the in vivo distribution of the mouse are highlighted. The traditional bone specimen is complex in manufacturing process and needs to be assembled with bones, the traditional transparent bone specimen is not convenient to display and carry because the traditional transparent bone specimen needs to be stored in glycerol, and the glycerol needs to be replaced regularly due to the color loss and the mildewing in the long-time storage process. Therefore, the existing bone specimen preparation technology of mice needs to be improved, so that the mouse bone specimen is beautiful, portable, convenient to show, scientific and educational. There is a need for a new method for solving the above-mentioned problems existing in mouse bone specimens.

Disclosure of Invention

Based on the technical problems in the background art, the invention provides the preparation method of the mouse double-dyed transparent skeleton plastic-packaged embedded specimen, which can show the distribution condition of hard bones and cartilages in a mouse body and the relationship between the mouse skeleton and muscles while keeping the integrity of a skeleton system, is quick in preparation method and beneficial to production, and the prepared specimen is convenient to store, transport and display.

The method for manufacturing the mouse double-dyed transparent skeleton plastic-package embedded specimen sequentially comprises the steps of material preparation, gradient ethanol dehydration and fixation, acetone degreasing, mixed staining agent dyeing, rehydration, pre-transparence, decoloration, gradient glycerol transparency treatment, plasticization and UV glue embedding, and finally the mouse double-dyed transparent skeleton plastic-package embedded specimen is obtained.

Preferably, the acetone gradient ethanol dehydration fixation step is as follows: and (3) soaking the mice subjected to the material preparation step in ethanol with the concentration gradient from low to high in sequence for dehydration and fixation treatment.

Preferably, the acetone degreasing step is as follows: taking out the mouse subjected to gradient ethanol dehydration fixation treatment from ethanol, soaking in anhydrous acetone, performing degreasing treatment, taking out the mouse from the anhydrous acetone after 20-28h, and soaking in water for 10-14 h.

Preferably, the step of dyeing with mixed dyes is as follows: soaking the mice degreased by acetone in a mixed dye solution for 20-28h, wherein the mixed dye solution is a mixture formed by sequentially mixing alizarin red stock solution, aliskiren blue stock solution, acetic acid and 75% ethanol solution.

Preferably, the volume ratio of the alizarin red stock solution to the alisnew blue stock solution to the acetic acid to the 75% ethanol solution is 1:0.8-1.2:0.8-1.2: 16-18.

Preferably, the alizarin red stock solution is a mixture formed by mixing 25mg of alizarin red and 25ml of 95% ethanol solution by volume, and the alisnew blue stock solution is a mixture formed by mixing 75mg of alisnew blue and 25ml of 95% ethanol solution by volume.

Preferably, the rehydration step comprises: and taking the mice subjected to the dyeing step of the mixed dyeing agent out of the mixed dyeing agent, and sequentially soaking the mice in absolute ethyl alcohol and pure water for 10-14 hours respectively.

Preferably, the pre-transparency step is: soaking the mice subjected to the rehydration step in a potassium hydroxide solution with the mass fraction of 2%, wherein the standard for stopping soaking is that the complete spines of the mice can be seen through semitransparent mouse muscles, the hard bones of the mice are light pink, and the cartilages of the mice are blue.

Preferably, the decoloring step is as follows: and (3) soaking the mice subjected to the pre-transparence step in a destaining solution I for 1-3 days, and then soaking the mice in a destaining solution II for 1-3 days.

Preferably, the transparent liquid I is a mixture of a potassium hydroxide solution with the mass fraction of 1% and pure glycerol which are uniformly mixed in a volume ratio of 1:0.8-1.2, and the transparent liquid II is a mixture of water and pure glycerol which are uniformly mixed in a volume ratio of 1: 0.8-1.2.

Preferably, the gradient glycerol transparent treatment is as follows: and (3) sequentially immersing the mice subjected to the decolorization step into glycerol with the volume fraction increasing gradually for 1-3 days respectively, and then immersing the mice into pure glycerol until the muscles are colorless and transparent and the bone structures are clearly visible.

Preferably, the plasticizing step is: selecting PEG-600 as a plasticizer, adding thymol crystals into a potassium hydroxide solution with the mass fraction of 0.5%, fully mixing, mixing the potassium hydroxide solution added with the thymol crystals and the PEG-600 according to a certain proportion to respectively prepare a mixture with the volume concentration of 15%, 25%, 50%, 75% and 100%, sequentially putting the mice into the mixture for soaking for 1-2d when the concentration is 15%, and then soaking for 2-3d when each gradient is followed, so that the glycerol in the PEG-600 can be completely replaced by the glycerol to finish plasticization.

Preferably, the step of embedding the UV glue comprises: pouring UV glue into a silica gel mold, stopping pouring when the UV glue does not cover 1cm of the bottom of the mold, placing the mold in an ultraviolet irradiation environment, placing a mouse subjected to a plasticizing step into the mold after the UV glue on the bottom layer is solidified, adjusting the posture, then pouring the UV glue until the mouse does not cover about 1cm of a specimen, placing the mold in the ultraviolet irradiation environment, demolding and taking out the embedded specimen after the UV glue is completely solidified, and polishing and finishing to obtain the mouse double-dyed transparent skeleton plastic package embedded specimen.

Compared with the prior art, the invention has the beneficial technical effects that:

(1) compared with the traditional mouse bone specimen, the specimen can not completely split the bone, so that the tedious process of reassembling bone parts is avoided, and the specimen manufacturing steps are saved. Moreover, because alizarin red can be specifically combined with calcium in bones, and alisnew blue can be combined with keratan sulfate in cartilage under the condition of proper pH, the two stains can obviously identify the distribution of hard bones and cartilage in mice, and the function is not possessed by traditional mouse bone specimens.

(2) Compared with the traditional transparent bone specimen, the specimen adopts the solution of plasticizing firstly and embedding secondly, and effectively solves the problems that the traditional transparent bone specimen is required to be stored in a container filled with glycerol, is not easy to carry and display and can be decolorized. The mouse transparent skeleton plasticized and embedded specimen manufactured by the method has the advantages of being transparent, plasticized and embedded, and is convenient to display, carry, store and transport.

Drawings

FIG. 1 is a schematic view of a mouse double-stained transparent skeleton plastic-encapsulated embedded specimen according to the present invention;

FIG. 2 is a schematic diagram of the skeletal structure of a mouse according to the present invention.

Detailed Description

The reagents involved in the present invention are commercially available.

Example 1

The invention provides a method for manufacturing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen, which comprises the following steps:

s1: preparing materials: collecting dead mice born for 4d, carefully peeling skin tissues of the mice by using an ophthalmologic forceps, avoiding damaging structures at the toe end and the tail part of the mice by mistake when peeling the skin, not peeling the skin at the tail end of four limbs, and removing fat tissues at the shoulder blade and the neck in advance for the integral aesthetic appearance of a specimen, but paying attention not to damage the integrity of the bones of the mice, then removing the internal organs of the mice, and washing the mice by using clear water;

s2: gradient ethanol dehydration fixation: soaking the mice subjected to the material preparation step into ethanol with concentration gradients of 80%, 90% and 100% in sequence for dehydration and fixation;

s3: acetone degreasing: taking out the mouse subjected to gradient ethanol dehydration fixation treatment from ethanol, soaking the mouse in anhydrous acetone, carrying out degreasing treatment, taking out the mouse from the anhydrous acetone after 24 hours, and soaking the mouse in water for 12 hours;

s4: dyeing with a mixed dyeing agent: soaking a mouse degreased by acetone in a mixed dye solution for 24 hours, wherein the mixed dye solution is a mixture formed by sequentially mixing 25mL of alizarin red stock solution, 25mL of aliskiren blue stock solution, 25mL of acetic acid and 425mL of ethanol solution with the volume concentration of 75%; the alizarin red stock solution is a mixture formed by mixing 25mg of alizarin red and 25ml of 95% ethanol solution in volume concentration, and the alisnew blue stock solution is a mixture formed by mixing 75mg of alisnew blue and 25ml of 95% ethanol solution in volume concentration;

s5: rehydration: taking the mice which are subjected to the dyeing step of the mixed dyeing agent out of the mixed dyeing agent, and sequentially soaking the mice in absolute ethyl alcohol and pure water for 12 hours respectively;

s6: pre-transparency: soaking the mice subjected to the rehydration step in a potassium hydroxide solution with the mass fraction of 2%, wherein the standard of stopping soaking is that the complete spines of the mice can be seen through semitransparent mouse muscles, the hard bones of the mice are light pink, and the cartilages of the mice are blue;

s7: and (3) decoloring: soaking the mice subjected to the pre-transparency step in a destaining solution I for 2d, and then soaking the mice in a destaining solution II for 2 d;

s8: gradient glycerol transparent treatment: sequentially immersing the mice subjected to the decoloring step into glycerol with gradually increased volume fraction for 2d respectively, and then immersing the mice into pure glycerol for soaking until the muscles are colorless and transparent and the bone structures are clearly visible; the transparent liquid I is a mixture formed by uniformly mixing 1% by mass of a potassium hydroxide solution and 1:1 by volume of pure glycerol, and the transparent liquid II is a mixture formed by uniformly mixing 1:1 by volume of water and pure glycerol;

s9: plasticizing: selecting PEG-600 as a plasticizer, adding thymol crystals into a potassium hydroxide solution with the mass fraction of 0.5%, fully mixing, mixing the potassium hydroxide solution added with the thymol crystals and the PEG-600 according to a certain proportion to respectively prepare a mixture with the volume concentration of 15%, 25%, 50%, 75% and 100%, sequentially putting the mice into the mixture for soaking for 2d when the concentration of the mice is 15%, and then soaking for 3d when each gradient is followed, so that the glycerol in the PEG-600 can be completely replaced by the PEG-600 to complete plasticization;

s10: and (5) embedding with UV glue: pouring UV glue into a silica gel mold, stopping pouring when the UV glue does not cross 1cm of the bottom of the mold, placing the mold in an ultraviolet irradiation environment, placing a mouse subjected to a plasticizing step into the mold after the UV glue on the bottom layer is solidified, adjusting the posture, then pouring the UV glue until the mouse does not cross about 1cm of a specimen, placing the mold in the ultraviolet irradiation environment, demolding and taking out the embedded specimen after the UV glue is completely solidified, polishing and finishing to obtain the mouse double-dyed transparent skeleton plastic package embedded specimen, wherein the silica gel mold is a cuboid silica gel mold (50 multiplied by 85 multiplied by 50, unit mm).

Example 2

The invention provides a method for manufacturing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen, which comprises the following steps:

s1: preparing materials: collecting dead mice born for 4d, carefully peeling skin tissues of the mice by using an ophthalmologic forceps, avoiding damaging structures at the toe end and the tail part of the mice by mistake when peeling the skin, not peeling the skin at the tail end of four limbs, and removing fat tissues at the shoulder blade and the neck in advance for the integral aesthetic appearance of a specimen, but paying attention not to damage the integrity of the bones of the mice, then removing the internal organs of the mice, and washing the mice by using clear water;

s2: gradient ethanol dehydration fixation: soaking the mice subjected to the material preparation step into ethanol with concentration gradients of 80%, 90% and 100% in sequence for dehydration and fixation;

s3: acetone degreasing: taking out the mouse subjected to gradient ethanol dehydration fixation treatment from ethanol, soaking the mouse in anhydrous acetone, carrying out degreasing treatment, taking out the mouse from the anhydrous acetone after 20 hours, and soaking the mouse in water for 10 hours;

s4: dyeing with a mixed dyeing agent: soaking a mouse degreased by acetone in a mixed dye solution for 20 hours, wherein the mixed dye solution is a mixture formed by sequentially mixing 25mL of alizarin red stock solution, 20mL of aliskiren blue stock solution, 20mL of acetic acid and 400mL of 75% ethanol solution with volume fraction; the alizarin red stock solution is a mixture formed by mixing 25mg of alizarin red and 25ml of 95% ethanol solution in volume concentration, and the alisnew blue stock solution is a mixture formed by mixing 75mg of alisnew blue and 25ml of 95% ethanol solution in volume concentration;

s5: rehydration: taking the mice which are subjected to the dyeing step of the mixed dyeing agent out of the mixed dyeing agent, and sequentially soaking the mice in absolute ethyl alcohol and pure water for 10 hours respectively;

s6: pre-transparency: soaking the mice subjected to the rehydration step in a potassium hydroxide solution with the mass fraction of 2%, wherein the standard of stopping soaking is that the complete spines of the mice can be seen through semitransparent mouse muscles, the hard bones of the mice are light pink, and the cartilages of the mice are blue;

s7: and (3) decoloring: soaking the mice subjected to the pre-transparence step in a destaining solution I for 1d, and then soaking the mice in a destaining solution II for 1 d;

s8: gradient glycerol transparent treatment: sequentially immersing the mice subjected to the decoloring step into glycerol with gradually increased volume fraction for 1d respectively, and then immersing the mice into pure glycerol for soaking until the muscles are colorless and transparent and the bone structures are clearly visible; the transparent liquid I is a mixture formed by uniformly mixing 1% by mass of a potassium hydroxide solution and pure glycerol in a volume ratio of 1:0.8, and the transparent liquid II is a mixture formed by uniformly mixing water and pure glycerol in a volume ratio of 1: 0.8;

s9: plasticizing: selecting PEG-600 as a plasticizer, adding thymol crystals into a potassium hydroxide solution with the mass fraction of 0.5%, fully mixing, mixing the potassium hydroxide solution added with the thymol crystals and the PEG-600 according to a certain proportion to respectively prepare a mixture with the volume concentration of 15%, 25%, 50%, 75% and 100%, sequentially putting the mice into the mixture for soaking for 1d when the concentration of the mice is 15%, and soaking for 2d when each subsequent gradient is formed, so that the glycerol in the PEG-600 can be completely replaced by the PEG-600 to complete plasticization;

s10: and (5) embedding with UV glue: pouring UV glue into a silica gel mold, stopping pouring when the UV glue does not exceed the bottom of the mold by 1cm, placing the mold in an ultraviolet irradiation environment, placing a mouse subjected to a plasticizing step into the mold after the bottom layer of the UV glue is solidified, adjusting the posture of the mouse, pouring the UV glue until the mouse does not exceed the specimen by about 1cm, placing the mold in the ultraviolet irradiation environment, demolding and taking out the embedded specimen after the UV glue is completely solidified, polishing and finishing to obtain the mouse double-dyed transparent skeleton plastic package embedded specimen, wherein the silica gel mold is a cuboid silica gel mold (50 multiplied by 85 multiplied by 50, unit mm)

Example 3

The invention provides a method for manufacturing a mouse double-dyed transparent skeleton plastic-packaged embedded specimen, which comprises the following steps:

s1: preparing materials: collecting dead mice born for 4d, carefully peeling skin tissues of the mice by using an ophthalmologic forceps, avoiding damaging structures at the toe end and the tail part of the mice by mistake when peeling the skin, not peeling the skin at the tail end of four limbs, and removing fat tissues at the shoulder blade and the neck in advance for the integral aesthetic appearance of a specimen, but paying attention not to damage the integrity of the bones of the mice, then removing the internal organs of the mice, and washing the mice by using clear water;

s2: gradient ethanol dehydration fixation: soaking the mice subjected to the material preparation step into ethanol with concentration gradients of 80%, 90% and 100% in sequence for dehydration and fixation;

s3: acetone degreasing: taking out the mice subjected to gradient ethanol dehydration fixation treatment from ethanol, soaking the mice in anhydrous acetone, carrying out degreasing treatment, taking out the mice from the anhydrous acetone after 28 hours, and soaking the mice in water for 14 hours;

s4: dyeing with a mixed dyeing agent: soaking a mouse degreased by acetone in a mixed dye solution for 28 hours, wherein the mixed dye solution is a mixture formed by sequentially mixing 25mL of alizarin red stock solution, 30mL of alisin blue stock solution, 30mL of acetic acid and 450mL of 75% ethanol solution with volume fraction; the alizarin red stock solution is a mixture formed by mixing 25mg of alizarin red and 25ml of 95% ethanol solution in volume concentration, and the alisnew blue stock solution is a mixture formed by mixing 75mg of alisnew blue and 25ml of 95% ethanol solution in volume concentration;

s5: rehydration: taking the mice which are subjected to the dyeing step of the mixed dyeing agent out of the mixed dyeing agent, and sequentially soaking the mice in absolute ethyl alcohol and pure water for 14 hours respectively;

s6: pre-transparency: soaking the mice subjected to the rehydration step in a potassium hydroxide solution with the mass fraction of 2%, wherein the standard of stopping soaking is that the complete spines of the mice can be seen through semitransparent mouse muscles, the hard bones of the mice are light pink, and the cartilages of the mice are blue;

s7: and (3) decoloring: soaking the mice subjected to the pre-transparency step in a destaining solution I for 3d, and then soaking the mice in a destaining solution II for 3 d;

s8: gradient glycerol transparent treatment: sequentially immersing the mice subjected to the decoloring step into glycerol with gradually increased volume fraction for 3d respectively, and then immersing the mice into pure glycerol until the muscles are colorless and transparent and the bone structures are clearly visible; the transparent liquid I is a mixture formed by uniformly mixing 1% by mass of a potassium hydroxide solution and 1:1.2 by volume of pure glycerol, and the transparent liquid II is a mixture formed by uniformly mixing 1:1.2 by volume of water and pure glycerol;

s9: plasticizing: selecting PEG-600 as a plasticizer, adding thymol crystals into a potassium hydroxide solution with the mass fraction of 0.5%, fully mixing, mixing the potassium hydroxide solution added with the thymol crystals and the PEG-600 according to a certain proportion to respectively prepare a mixture with the volume concentration of 15%, 25%, 50%, 75% and 100%, sequentially putting the mice into the mixture for soaking for 2d when the concentration of the mice is 15%, and then soaking for 3d when each gradient is followed, so that the glycerol in the PEG-600 can be completely replaced by the PEG-600 to complete plasticization;

s10: and (5) embedding with UV glue: pouring UV glue into a silica gel mold, stopping pouring when the UV glue does not exceed the bottom of the mold by 1cm, placing the mold in an ultraviolet irradiation environment, placing a mouse subjected to a plasticizing step into the mold after the bottom layer of the UV glue is solidified, adjusting the posture of the mouse, pouring the UV glue until the mouse does not exceed the specimen by about 1cm, placing the mold in the ultraviolet irradiation environment, demolding and taking out the embedded specimen after the UV glue is completely solidified, polishing and finishing to obtain the mouse double-dyed transparent skeleton plastic package embedded specimen, wherein the silica gel mold is a cuboid silica gel mold (50 multiplied by 85 multiplied by 50, unit mm)

Fig. 1-2 show a double-stained transparent skeleton plastic-encapsulated mouse bone specimen obtained by the method of example 1, which ensures the integrity of mouse skeleton and significantly identifies the distribution of hard bone and cartilage of mouse.

The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.