A mounting medium for sealing blood smears and a method for mounting the same

Abstract

The present invention relates to the technical field of biochemical experiments, and in particular to a mounting medium for sealing blood smears and a method for mounting the same. Characterized by volume parts, the mounting medium comprises 3 parts of bisphenol A epoxy resin, 1 part of diethylenetriamine and 1 part of photocurable acrylate resin. After mixing the components of the mounting medium evenly, let it stand for 3 to 5 minutes until the bubbles disappear, and then use it. Place the applicator on one side of the blood film on the glass slide and slowly translate it to the other side to coat the mounting medium into a liquid film with a thickness of 0.1 to 0.5 mm and completely covering the blood film. Let it stand for 10 minutes, irradiate it with a 6W 360-395 nm ultraviolet lamp for 3 minutes, and then move the blood smear into a dust-proof cover and let it stand at room temperature for 20 to 24 hours. This method of machine mounting the mounting medium increases the service life of the blood smear, facilitates permanent preservation, and has broad application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of biochemical experiments, and in particular to a mounting medium for sealing blood smears and a method for mounting the same. Background Art

[0002] A blood smear is to smear a blood specimen on a glass slide, and after staining, the cell morphology and changes are examined under a microscope. It is a basic method for clinical blood cytology examination and is widely used, and has great diagnostic value for various blood diseases. The blood film of the blood smear is exposed outside and is prone to mildew and insect damage. Especially after the blood smear is examined with an oil immersion lens, a deoiling agent such as xylene is required to remove the lens oil on the blood film, which is extremely likely to cause the blood smear to fade, be contaminated and damaged, and is not conducive to the reexamination of the blood smear. Especially for the blood smear used for teaching demonstration, the blood film is easily scratched and seriously contaminated, resulting in a short service life. Therefore, it is necessary to mount the blood smear. The conventional mounting method is to store the blood sample between the glass slide and the coverslip, and bond them through the glass slide and the coverslip to complete the sealing of the blood sample. Since the mounting link avoids the further oxidation and color degradation of the blood sample, the storage time of the blood specimen sealed with the mounting medium is extended. Based on the use of the mounting medium, the current performance requirements for the mounting medium are: 1. The refractive index is close to that of the glass slide or the coverslip; 2. The connection between the coverslip and the glass slide can be maintained relatively firmly; 3. There is no interaction with the stain of the specimen. According to these characteristics, the commonly used mounting media in experiments are mainly divided into the following two categories: 1. Non-water-soluble mounting media represented by neutral gum and Canada balsam. After staining, the specimen slide is sealed after dehydration with gradient alcohol and further transparency with xylene. 2. Water-soluble mounting media represented by glycerin gelatin and arabic gelatin.

[0003] However, in actual operation, it is easy to crush the coverslip on the glass slide when the objective lens moves down. Therefore, technicians gradually try to seal the blood smear by directly covering the mounting medium on the blood sample. This method needs to ensure that the colloid will not change the cell morphology, the transparency of the colloid itself, and the stability of long-term storage. If the conventional neutral gum and Canada balsam are directly used to seal the blood sample on the glass slide, the curing time is usually more than one week, and both neutral gum and Canada balsam have certain requirements in use and storage. Most importantly, both need to be used in combination with the solvent xylene or benzene, and these solvents will affect the health of the users. In addition, the surface is uneven after applying the neutral gum and Canada balsam. If glycerin gelatin and arabic gelatin are directly used to seal the blood sample, it is necessary to heat and dissolve the colloid first, and the high temperature will cause the cells to deform, and the colloid is easy to turn yellow and brittle after curing, which is not suitable for directly sealing the blood sample on the glass slide.

[0004] Publication No. 109749634 discloses a rapid mounting medium based on photocuring and its preparation method. The photocuring technology used can rapidly transform the polymer from a liquid into a high-performance solid coating. Since there is no dissolution and volatilization process, it is recognized as an environmentally friendly technology and has good adhesion on glass. However, if the photocuring monomer is directly placed on the blood sample on the glass slide, due to the too-fast curing speed and large shrinkage rate of the photocuring monomer, it is easy to cause deformation of the blood film and cells, and obvious astigmatism occurs during microscopic examination. Summary of the Invention

[0005] Aiming at the problem that the existing mounting media and mounting methods are not suitable for directly sealing blood smear samples on glass slides, the present invention proposes a mounting medium for sealing blood smears and its mounting method.

[0006] A mounting medium for sealing blood smears according to the present invention is characterized in that by volume, the mounting medium comprises 3 parts of bisphenol A epoxy resin, 1 part of diethylenetriamine and 1 part of photocuring acrylate resin, and is used after being mixed evenly.

[0007] Furthermore, the mounting medium further comprises polyether-modified silicone. By volume, the addition amount of polyether-modified silicone is 0.01 - 0.05 parts. Since bubbles are generated during the stirring and mixing process of the mounting medium, and the vacuum defoaming method can only eliminate the bubbles visible to the naked eye, there are still fine bubbles under the microscope; in addition, since the mounting medium realizes the curing process through a polymerization reaction, this process is an exothermic reaction, the fine bubbles expand when heated, the gas is no longer compatible with the epoxy system, and the generated bubbles will seriously interfere with the observation of cell morphology under the microscope. In the use of conventional epoxy resin adhesives, they can be placed in hot water for water-bath defoaming, but since the blood sample will cause morphological changes under the microscope after being heated, therefore, due to the fast defoaming speed and low surface tension of polyether-modified silicone, the mounting medium after adding polyether-modified silicone is easy to spread on the blood film surface, and the light transmittance for microscopic observation is relatively high.

[0008] Furthermore, the mounting medium also includes phthalate. By volume, the addition amount of phthalate is 0.1 - 0.5 parts. Phthalate can be well compatible with the mounting medium, can reduce the viscosity of the mounting medium polymer, and avoid blood film deformation caused by too large shrinkage rate.

[0009] Furthermore, the mounting medium also includes photoinitiator TPO. By volume, the addition amount of TPO is 0.03 - 0.1 parts, which is used to generate free radicals after receiving ultraviolet light, thereby triggering rapid curing of the mounting medium.

[0010] Further, the mounting medium further includes isocyanate. By volume, the addition amount of isocyanate is 0.05 - 0.4 parts, which can reduce the moisture permeability of the mounting medium and increase the water resistance, facilitating long-term preservation. In addition, since the curing method of the mounting medium of the present invention is a photocuring scheme, and bisphenol A epoxy resin is prone to yellowing after long-term preservation under ultraviolet light irradiation. After experiments, it is found that after adding isocyanate, in the ultraviolet resistance and aging tests, the mounting medium samples added with isocyanate do not turn yellow.

[0011] The method for mounting a blood smear of the present invention is characterized in that it is implemented by the following method: After mixing the components of the mounting medium evenly, let it stand for 3 - 5 minutes until the bubbles disappear, then use a glue applicator on one side of the blood film on the glass slide and slowly translate it to the other side to coat the mounting medium into a liquid film with a thickness of 0.1 - 0.5 mm that completely covers the blood film. Let it stand for 10 minutes, irradiate it with a 6W 360 - 395 nm ultraviolet lamp for 3 minutes, and then move the blood smear into a dust-proof cover and let it stand at room temperature for 20 - 24 hours.

[0012] Further, the structure of the glue applicator consists of a funnel and a rolling shaft. The opening at the bottom of the funnel is in a long strip shape, and buckles are provided on both sides of the opening. A rolling shaft matching the shape of the opening is fixed inside the buckles; protrusions are provided on both sides of the position of the opening at the bottom of the funnel, and the height of the protrusions is 0.1 - 0.5 mm.

[0013] When in use, put the evenly mixed mounting medium into the funnel. There is a certain gap between the rolling shaft and the opening. When rolling, the mounting medium evenly covers the surface of the rolling shaft and is transferred to the blood film to complete the mounting work. The protrusions can ensure that the rolling shaft will not squeeze the blood film during the moving process and ensure the integrity of the blood smear.

[0014] The mounting medium prepared by the present invention has medium viscosity, is colorless and transparent, non-toxic and odorless, does not generate heat, has a long operation time, a short curing time, strong adhesion, good flatness, low shrinkage rate, high hardness after curing, is insoluble in xylene, ethanol and water, and has good tolerance to high and low temperatures. After mounting, it sets quickly, has a flat surface, beautiful appearance, the blood film does not shrink, does not change color, has a clear field of view, no refraction and astigmatism phenomenon, does not mildew, is waterproof and mothproof, is easy to clean after use, has good wear resistance, increases the service life of the blood smear, and is convenient for permanent preservation. The glue applicator has a simple design, is easy to use, has uniform coating, and is convenient for mounting a large number of blood smears. This mounting method replaces the traditional method of using a cover glass combined with a mounting medium to mount blood smears, thus overcoming the technical defect of crushing the cover glass when the objective lens drops. The blood smear is clearly observed under the microscope and has a long preservation time, with broad application prospects. Description of the Drawings

[0015] Figure 1 It is a schematic structural diagram of the glue applicator.

[0016] Figure 2 It is a schematic bottom surface structural diagram of the glue applicator.

[0017] Among them, there are a funnel 1, a rolling shaft 2, and a protrusion 3. DETAILED DESCRIPTION

[0018] Example 1: The sealing agent used in this example is: by volume, 3 parts of bisphenol A epoxy resin, 1 part of diethylenetriamine and 1 part of light-curing acrylate resin, 0.05 parts of polyether-modified silicon, 0.5 parts of phthalate, 0.1 parts of TPO, and 0.4 parts of isocyanate are fully mixed and allowed to stand for 3 to 5 minutes for use. Use a glue applicator to one side of the blood film on the slide, slowly move it to the other side, and apply the sealing agent to a 0.1 to 0.5 mm thick liquid film that completely covers the blood film. Let it stand for 10 minutes, irradiate it with a 6W 360-395nm ultraviolet lamp for 3 minutes, and then move the blood smear into a dust cover and let it stand at room temperature for 20-24 hours.

[0019] Comparative Example: The mounting agent used in the comparative example is: by volume, 2 parts of bisphenol A epoxy resin, 1 part of diethylenetriamine and 1 part of light-curing acrylate resin, which are fully mixed and left to stand for 3 to 5 minutes. The mounting agent of the comparative example is applied to the slide glass using a glue applicator, and other experimental conditions remain unchanged. The surface of the blood smear prepared by the mounting agent of this component is not smooth and has a low hardness. Although there are no obvious bubbles in naked eye observation, there are many bubbles under microscopy, which will interfere with the microscopic observation.

Claims

1. A mounting medium for sealing blood smears, characterized in that by volume, the mounting medium comprises 3 parts of bisphenol A epoxy resin, 1 part of diethylenetriamine and 1 part of photocurable acrylate resin, which are mixed evenly before use; the mounting medium also includes phthalate, by volume, the addition amount of phthalate is 0.1 - 0.5 parts, and phthalate is well compatible with the mounting medium, reducing the viscosity of the mounting medium polymer and avoiding blood film deformation caused by excessive shrinkage rate.

2. A mounting medium for sealing blood smears according to claim 1, characterized in that the mounting medium further comprises polyether-modified silicone, by volume, the addition amount of polyether-modified silicone is 0.01 - 0.05 parts.

3. A mounting medium for sealing blood smears according to claim 1, characterized in that the mounting medium also includes photoinitiator TPO, by volume, the addition amount of TPO is 0.03 - 0.1 parts; the mounting medium also includes isocyanate, by volume, the addition amount of isocyanate is 0.05 - 0.4 parts.

4. A method for mounting a blood smear using the mounting medium for sealing blood smears according to claim 1, characterized in that it is implemented by the following method: after mixing the components of the mounting medium evenly, let it stand for 3 - 5 minutes until the bubbles disappear, then use it. Place the applicator on one side of the blood film on the glass slide and slowly translate it to the other side to coat the mounting medium into a liquid film with a thickness of 0.1 - 0.5 mm that completely covers the blood film. Let it stand for 10 minutes, irradiate it with a 6W 360 - 395 nm ultraviolet lamp for 3 minutes, and then move the blood smear into a dust-proof cover and let it stand at room temperature for 20 - 24 hours.

5. A method for mounting a blood smear using the mounting medium for sealing blood smears according to claim 4, characterized in that the structure of the applicator consists of a funnel and a rolling shaft. The opening at the bottom of the funnel is rectangular, and clips are provided on both sides of the opening. A rolling shaft matching the shape of the opening is fixed inside the clips; protrusions are provided on both sides of the position of the opening at the bottom of the funnel, and the height of the protrusions is 0.1 - 0.5 mm.

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