KR100752420B1 - Manufacture of biological specimen and manufactured biological specimen using thereof - Google Patents

Abstract

A biological specimen manufacturing method and a biological specimen produced by the same are provided to permanently preserve the biological specimen by fixing the biological specimen through acryl resin and sterilizing the biological specimen through radiant rays. A biological specimen manufacturing method is composed of a step of pouring the acryl resin mixed aqueous solution and a biological specimen into a container and then closing up the container and a step of irradiating radiant rays to the closed container. The acryl resin is at least two selected from a group comprising acrylamide, bisacrylamide, acrylonitrile, methylmethacrylate, and styrene monomer. In particular, the acrylamide and the bisacrylamide are mixed. The acrylamide of 3~20g is dissolved in the water of 100ml.

Description

Manufacture method of biological specimen and manufactured biological specimen using same

1 is a biological specimen (a: carnation, b: freesia, c: gypsophila, d: pussy, e: carnation) prepared by curing acrylamide by irradiating gamma rays after putting an aqueous acrylamide solution and a plant specimen in a transparent container. Indicates.

The present invention relates to a method for producing a biological sample and to a biological sample produced using the same.

Existing biological specimens are manufactured by the method of drying, stuffing, immersing, curing with synthetic resin, etc., which are used for drying materials such as plants, animals, and insects. We use for purpose. The method of manufacturing such a specimen is selected according to the type and characteristics of the organism to be sampled and the purpose of use. Biological specimens are largely divided into plants, insects, and animals according to the types of organisms, and include microorganisms and tissues for microscopic observation. Conventionally used methods for producing a biological sample include a method of large drying, a method of stuffing, a liquid immersion method and a method of curing with a synthetic resin.

First, the drying method is one of the most common methods for producing biological specimens, and is mainly used for preparing specimens of plants and insects, such as seaweeds, herbs and wood. Sampling of plants generally involves taking plants, placing them between papers, such as newspapers, fixing them, and replacing them with new paper for about a week to remove moisture. After removing the moisture, fix it on a fixed plate of the appropriate size and complete the specimen by writing the name of the plant, collecting place, and collector. Sampling of insects is made mainly by inserting pins into collected insects, then making the shape of wings, legs, etc. in a frame to fix the appearance, and drying them in air. Dry specimens have the advantage that they can be produced relatively easily without special equipment. However, dry specimens have problems such as discoloration and decay in the manufacturing process and storage process, and in the case of plant specimens, it is difficult to preserve the original form and takes a long time to produce. In particular, the dry specimen is a state in which water is removed, but there is a problem in that it is difficult to preserve it for a long time due to frequent decay by microorganisms such as molds and loss by micro insects such as ants during the preservation process.

Next, the stuffing method is a sampling method of large animals with a large amount of water, such as birds and mammals. The manufacturing process of the stuffing consists of removing the parts with high water content such as the internal organs and muscles of the animal, preserving them, and then filling the space with non-corrosive materials and drying them to maintain the appearance. The production of the stuffed requires skill and the cost is very high.

Next, the immersion method is a sample manufacturing technique applied to the case where the moisture content of plants, animal organs, molluscs, etc. is high, and shape preservation is difficult by drying. The manufacturing method consists of putting the sample in a glass or plastic bottle, pouring a preservative, and then sealing it. The preservatives vary depending on the type of organism and the purpose of sample preparation, but generally 40 to 80% of ethanol or 20 to 40% of formalin is commonly used, and in some cases, a solution added with an acid such as acetic acid may be used. The immersion specimen has the advantage of keeping the original form of the organism unlike the dry specimen, but it is large in fluidity because it is a form in which the sample is soaked in an aqueous solution. have.

In order to solve these problems of the above-mentioned preservation methods, a method of preparing a biosample by curing a resin was devised. The method for producing a biological sample by curing the synthetic resin is a manufacturing method for fixing the biological sample to the transparent resin by fixing the biological sample to a synthetic resin solution such as polyester and heating the same to induce curing of the resin. Although these methods have improved aspects of preservation and ease of use of biological specimens, they still have complex manufacturing methods, require a long time of about a week, require a large amount of chemicals, and are difficult to apply to large biological specimens.

As discussed above, biological specimens are of great value as commodities because of the growing demand for not only educational specimens but also various souvenirs and decorations. However, the conventional methods for making specimens such as drying, stuffing, immersion, synthetic resin curing, and the specimens produced therefrom are limited in their fabrication methods, difficult to preserve their original appearance, limited in long-term preservation, and There are problems such as manufacturing period and high manufacturing cost. Therefore, it is necessary to maintain or improve the quality characteristics inherent in biological specimens, to apply them regardless of the type or size of the organisms, and to develop new biological specimens that are not only simple in manufacturing but also semi-permanent for long-term preservation. Do.

Irradiation technology is a sterilization technology certified by various international organizations (FAO, WHO, IAEA, ICGFI, FDA, etc.) and is currently used for sterilization of food and public health products in more than 50 countries. It is a technology that is widely used in the polymer industry using the property of inducing.

Such radiation can expect selective sterilization effect while minimizing changes in the quality of the target product, and does not generate heat and can be sterilized in a completely packaged state, thus preventing secondary pollution during the packaging process. have. In addition, radiation can induce a polymerization reaction such as a synthetic resin without chemical reaction additives, it is possible to produce a variety of polymer materials and improve the properties of the material. By utilizing these characteristics of radiation, various technologies and products are being developed in the fields of hydrogel and special polymer products worldwide.

There are no examples of the application of technology to improve the convenience of biosampling and increase the product quality and added value using radiation sterilization and radiation curing technology.

Accordingly, the present inventors endeavored to solve the problems of the method of drying, stuffing, immersion and curing with a synthetic resin, and as a result, put the biological specimen in a container containing an acrylic resin mixed aqueous solution such as acrylamide, and then After sealing, it was confirmed that the microorganisms inhabiting the biological specimens were sterilized while being irradiated with radiation to induce solidification or curing of an acrylic resin mixed aqueous solution such as acrylamide. Thus, the present inventors have completed the present invention by confirming that the preservation properties are improved while the characteristics originally possessed by the biological specimen prepared by the above-described manufacturing method are not changed.

The present invention maintains or improves the inherent quality characteristics of biological specimens, and can be applied regardless of the type or size of the organism. And it is an object to provide a biological specimen prepared using the same.

The present invention comprises putting the acrylic resin mixed aqueous solution and the biosample in a suitable container and then sealing it (step 1); And it provides a method for producing a semi-permanent preservation possible biological specimens comprising the step (step 2) of irradiating the sealed container and a biological specimen prepared using the same.

The acrylic resin mixed aqueous solution of step 1 according to the present invention may be used an acrylic monomer. In this case, the acryl-based resin may be acrylamide, bisacrylamide, acrylonitrile, methyl methacrylate, styrene monomer, and the like. Preferably, acrylamide and bisacrylamide may be used.

The acrylamide and bis acrylamide can be dissolved in an aqueous solution, and acrylamide is preferably dissolved in 3 g to 20 g with respect to 100 ml of water. The acrylamide concentration range is not only a range that can be easily cured by radiation, but when the acrylamide is less than 3 g, when it is cured, fluidity is difficult to support the biosample, and when it is more than 20 g It is not preferable because curvature is observed in the cured acrylamide.

An aqueous solution in which the acrylic monomer is dissolved in water is generally used, but the type of the synthetic resin mentioned above is just one example, and the content of the present invention is not limited thereto. Acrylic monomers are liquid plastics having water resistance, acid resistance, alkali resistance and oil resistance, and when the monomers are polymerized and cured, the light transmittance is superior to glass.

In addition, the sample that can be used in step 1 is not particularly limited, and preferably, plants, insects, animals and parts or organs thereof which are biosamples can be used. When the specimen is dried to prepare the specimen, it is difficult to preserve the original shape of the organism as it is, and it takes a long time to produce, and is likely to be damaged by mold or insects. In case of stuffing, it is difficult to make a sample because it requires special skills. In addition, the immersion can maintain the inherent form of the organism, but the discoloration by the sample solution appears, there is a disadvantage that the movement of the sample is not easy to be fixed because it is not fixed. Finally, the synthetic resin curing method by chemical treatment and heating has a problem that the process is complicated, takes a long time and the size of the sample is limited.

Next, step 2 may induce curing of acrylamide by irradiating a sealed container containing a biosample and acrylamide. The radiation may be alpha rays, beta rays or gamma rays, it is preferable to use gamma rays.

Gamma rays are widely used in the medical field for the treatment of cancer, and are useful for modifying the internal bonds of metal materials or polymer materials. In the present invention, the water-soluble acrylic resin may induce polymer formation by irradiating the water-soluble acrylic resin without the introduction of other chemical additives, thereby inducing curing of the acrylic resin. The gamma radiation dose range in step 1 is 0.5 to 50 k 0.5, preferably 3 to 10 k㏉. The gamma-irradiation dose is a range in which curing of the acrylic resin mixed aqueous solution can be easily performed, and sterilization of microorganisms can be easily made in the above range.

In addition, the source of the gamma ray is not particularly limited, but preferably cobalt-60 may be used as the source.

Biosamples prepared according to the present invention can be sterilized by irradiation to completely remove microorganisms.

The quality of biological specimens is important for the reproducibility of their inherent appearance and the preservation to maintain them for a long time. In particular, since organisms inhabit the microorganisms derived from the natural state, corruption by the microorganisms becomes a problem. In order to prevent the decay of the organisms, the biological specimens are washed or treated with other chemicals, but the microorganisms are not completely removed from the biological specimens produced by the washing or other chemical treatment methods, and the biological specimens are removed from the air. If exposed, they can be easily contaminated with microorganisms.

Radiation can be expected to be selective sterilization effect while minimizing changes in the quality of the object, heat is not generated during irradiation, sterilization after sealing is possible there is no possibility of secondary pollution.

In addition, the manufacturing method of the biological sample is fixed by inducing the polymerization reactant of acrylamide by radiation has the advantage that can compensate for all the disadvantages of the dry sample, the stuffed sample, the immersion sample and the synthetic resin cured sample. In particular, sterilization and curing of biological specimens by radiation can be processed in a single process within hours without additional additives or pretreatment, thereby reducing process costs. In addition, the production of biological specimens by radiation can be immediately applied to all existing transparent containers including plastic and glass without making a separate container, and various types of products can be produced because they are not dependent on the type or characteristics of living organisms.

In addition, the manufacturing method according to the present invention may further comprise the step of removing bubbles from the acrylic resin mixed aqueous solution and the biological sample. In order to prevent the generation of air bubbles around the biological specimen and transparency in the product during the production of the biological specimen, it can be used to suppress the generation of bubbles by immersing in ice water, and remove the bubbles present in the acrylamide aqueous solution In order to remove the air bubbles in the mixed aqueous solution, a vacuum reducer can be used.

The present invention provides a biological specimen capable of semi-permanent preservation prepared using the method for producing a biological sample.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, the following examples are merely to illustrate the present invention, but the content of the present invention is not limited by the following examples.

Preparation Example 1 Preparation of Acrylamide-Bisacrylamide Mixed Solution

1-1: an aqueous solution of 3 g of acrylamide and bisacrylamide in 100 ml of water

In order to set the appropriate curing concentration of the curing acrylic resin to fix the sample, the mixed aqueous solution of acrylamide and bisacrylamide was completely dissolved in 3 g of acrylamide with respect to 100 ml of water. The mixed aqueous solution was treated for 5 minutes in a vacuum reducer to remove bubbles. The irradiation amount of gamma rays was 3 or 5 kPa to induce curing of the mixed aqueous solution, and the results are shown in Table 1 below.

1-2 to 1-7: other acrylamide-bisacrylamide aqueous solution

Acrylamide in the acrylamide-bisacrylamide mixed aqueous solution is the same method as in Preparation Example 1-1, except that 5 g, 7.5 g, 10 g, 15 g, 20 g and 30 g are used with respect to 100 ml of water. 5 g, 7.5 g, 10 g, 15 g, 20 g and 30 g of cured acrylamide were prepared by inducing curing of the mixed aqueous solution.

Component ratios of Preparation Examples 1-1 to 1-7 are shown in Table 1 below.

Mixing ratio of acrylamide-bisacrylamide aqueous solution for biosampling Water (ml) Acrylamide (g) Bisacrylamide (g) Preparation Example 1-1 100 3 0.1 Preparation Example 1-2 100 5 0.17 Preparation Example 1-3 100 7.5 0.25 Preparation Example 1-4 100 10 0.33 Preparation Example 1-5 100 15 0.5 Preparation Example 1-6 100 20 0.67 Preparation Example 1-7 100 30 One

Although all samples were cured according to the preparation example, some fluidity was observed in 3 g of acrylamide in 100 ml of water, and some bending was observed in more than 20 g. Thus, it was determined that 5 g to 20 g acrylamide was suitable for the production of radiation cured biosamples. In addition, the gamma-irradiation dose 3 or 5 kPa induced hardening without any influence on hardening of the mixed aqueous solution.

Example 1 Preparation of Carnation Specimens

The carnation was soaked in ice water for about 1 minute to prevent bubbles from being generated when the acrylamide aqueous solution was added. The carnation was cut to a length of about 5 cm to fit the sample vessel and then properly placed in the vessel. The acrylamide-bisacrylamide mixed aqueous solution of Preparation Example 4 was added to the vessel to immerse the carnation. After removing the microbubble in the mixed aqueous solution for 5 minutes in a vacuum reducer, the lid of the specimen container was immediately sealed. Carnation specimens were prepared by irradiating gamma rays at 3 k㏉ to the sealed specimen containers.

<Examples 2 to 4> Preparation of freesia, gypsophila and pussy

Freesia, gypsophila and pussy will be prepared in the same manner as in Example 1-1, except that freesia, gypsophila and pussy will each be used as biological specimens.

Example 5 Preparation of Carnation Specimens

Carnation samples were prepared in the same manner as in Example 1-1, except that the carnation samples were irradiated with gamma rays at 5 k㏉.

<Examples 6 to 8> Preparation of freesia, gypsophila and pussy

Freesia, gypsophila and pussy will be prepared in the same manner as in Example 1-5, except that freesia, gypsophila and pussy will be used as the biosamples, respectively.

<Comparative Examples 1 to 4> Biological specimens without irradiation of gamma rays

A gamma-irradiated carnation, freesia, gypsophila, and pussy specimens were prepared in the same manner as in Example 1-1, except that gamma-rays were not irradiated, and carnation, freesia, gypsophila, and willow, respectively.

Experimental Example 1 Sterilization and Preservation of Biological Specimens

The following experiments were performed to determine the microbial contamination of the biosamples irradiated with gamma rays and the contamination of the biological specimens without irradiated gamma rays.

10 g of the biological samples of Examples 1 to 8 and Comparative Examples 1 to 4 were diluted in 100 ml of sterile saline. Sterile saline diluted with the biological sample was stirred for 30 minutes at 4 ℃, and then diluted again several times in 1/10 unit, smeared on nutrient agar (Difco) medium and potato glucose agar (potato dextrose agar) medium It was. The nutrient agar medium and potato glucose agar medium were incubated in a microbial incubator at 30 ° C. for 2 days and the resulting microbial colonies were counted as the number of microorganisms per gram of biosample. In the nutrient agar medium, a colony of ordinary bacteria was formed, and in the potato glucose agar medium, mold formed a colony. The population of the common bacteria and fungi was counted and the results are shown in Table 2 below.

In addition, after preserving the biological specimens of carnations, freesia, gypsophila and pussy willows of Examples 5 to 8 irradiated with gamma rays at 5 k㏉ for 6 months at 30 ° C., the growth of microorganisms was observed, and the results are shown in FIG. 1. .

Bactericidal Effect According to Microbial Distribution and Gamma Irradiation General bacteria (colon count / g) Mold (colon / g) Example One 2.38 × 10 ² - 2 - - 3 1.21 × 10 ² 2.06 × 10 ¹ 4 8.44 × 10 ¹ - 5 - - 6 - - 7 - - 8 - - Comparative example One 4.86 × 10 ¹ 7.03 × 10 ³ 2 2.06 × 10 ³ 3.81 × 10 ¹ 3 5.44 × 10 ⁵ 4.65 × 10 ³ 4 4.92 × 10 ³ 5.72 × 10 ¹

As shown in Table 1, Examples 1 to 4 irradiated with gamma rays at 3 k㏉ significantly reduced the number of common bacteria and molds than Comparative Examples 1 to 4, whereas Examples 5 to 8 irradiated with gamma rays at 5 k㏉ are No colonies of common bacteria and fungi were formed. As a result, when the gamma ray was irradiated at 5 k㏉, the bactericidal effect against general bacteria and fungi was clearly seen.

In addition, as shown in Figure 1, Examples 5 to 8, which are biological specimens irradiated with gamma rays at 5 k 보존 were preserved for 30 months at 30 ℃ no general bacteria and fungi were observed. This confirmed that there is no risk of corruption by the microorganisms even if the biosamples of Examples 5 to 8 are stored for a long time.

The biosample according to the method of the present invention is fixed by the solidified acrylic resin and at the same time, the biosamples are sterilized by the irradiated radiation, thereby enabling permanent preservation of the biosample. In addition, not only the continuous processing of the manufacturing method is possible but also a process such as drying of a biological specimen can be omitted, thereby reducing the process cost. In addition, the method of manufacturing a biological specimen can be used in the production of learning materials, exhibitions, decorations, souvenirs and the like because it is possible to produce a variety of products by size and use.

Claims (12)

Putting the acrylic resin mixed aqueous solution and the biosample in a suitable container and then sealing it (step 1); And Method for producing a semi-permanent preservation, including the step of irradiating the sealed container (step 2). According to claim 1, wherein the acrylic resin of step 1 is at least two selected from the group consisting of acrylamide, bisacrylamide, acrylamide, bisacrylamide, acrylonitrile, methyl methacrylate and styrene monomer. Method of producing a biological sample. 3. The method of claim 2, wherein the acrylic resin mixture is a mixture of acrylamide and bisacrylamide. The method of claim 3, wherein the acrylamide is dissolved in 3 g to 20 g based on 100 ml of water. The method of claim 1, wherein the radiation of step 2 is alpha, beta, or gamma rays. The method of claim 5, wherein the radiation is gamma rays. The method of manufacturing a biological sample according to claim 5 or 6, wherein the irradiation dose of the radiation is 0.5 to 50 kPa. 8. The method of claim 7, wherein the radiation dose is 3 ~ 10 k㏉. The method of claim 1, wherein the biological sample is a plant, insect, animal, or part thereof. The method of claim 1, further comprising removing bubbles from the acrylic resin mixed aqueous solution or the biosample in step 1. The method of claim 10, wherein the bubble is removed by immersing the acrylic resin mixed aqueous solution or the biological sample in ice water or vacuum decompression. A biological specimen capable of semi-permanent preservation produced by the manufacturing method of claim 1.

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