Portable cervical exfoliated cell dyeing set and use method thereof

Abstract

The invention provides a portable cervical exfoliated cell staining suit and a use method thereof, and belongs to the technical field of cell staining devices. The portable cervical exfoliated cell dyeing suit comprises a bearing table, an elution device and a cover plate, a mixture 1 is attached to the bearing table, the elution device contains eluent, and a mixture 2 is attached to the cover plate; the mixture 1 is prepared from agarose, hematoxylin liquid and water; the eluent is composed of an ethanol solution, eosin dye liquor and water; the mixture 2 is composed of neutral gum and environment-friendly transparent liquid. By adopting the portable cervical exfoliated cell dyeing suit and matching with a corresponding specific dyeing treatment process, cervical exfoliated cell dyeing can be carried out in time when a gynecologic doctor examines, and the examination result time is effectively shortened.

Description

Technical Field

[0001] This invention belongs to the field of cell staining device technology, and particularly relates to a portable cervical exfoliated cell staining kit and its usage method. Background Technology

[0002] Cervical cancer is a common malignant tumor with a high incidence and mortality rate, posing a significant threat to women's health. Cervical cancer often has a precancerous lesion stage of about 10 years before diagnosis. Therefore, regular screening, guidance and referral for those who test positive in the initial screening, further diagnosis, and effective standardized disease management are extremely important for the early prevention and treatment of cervical cancer. my country also considers regular cervical cancer screening an important part of protecting women's health. Due to my country's vast territory and uneven economic development, in vast rural and remote areas, cervical smear tests are considered the main method of regular cervical cancer screening due to their low cost, and have significant clinical importance.

[0003] After performing a cervical smear examination, clinicians need to stain the smear and then use a microscope to diagnose the presence of tumor cells to assess the condition of the cervix. Currently, routine staining processes are commonly used for cervical smears (such as...). Figure 1 ) or fully automatic dyeing machine processing (such as Figure 2 However, both staining methods are space-consuming and inconvenient to carry. Currently, cervical smear tests require the collection of cervical smears from all patients during gynecological examinations, which are then sent to the pathology department for staining. The final diagnosis is made by a pathologist using a microscope, resulting in a delay of 1-2 days after the gynecologist's examination. Furthermore, the current cervical exfoliative cell staining method cannot be performed simultaneously with the gynecologist's examination, delaying timely clinical treatment for positive cases. A small number of positive cases are forgotten due to the long interval between the results and the examination, ultimately hindering their treatment. Therefore, there is an urgent need to develop a portable cervical exfoliative cell staining kit to shorten the examination time and ensure that positive cases receive timely clinical treatment. Summary of the Invention

[0004] In view of this, the purpose of this invention is to provide a portable cervical exfoliated cell staining kit and its method of use. This invention, by making the cervical exfoliated cell staining method portable and providing a corresponding specific staining process, enables timely cervical exfoliated cell staining during gynecological examinations, effectively shortening the time required for examination results.

[0005] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a portable cervical exfoliated cell staining kit, which includes a support platform, an elution device, and a cover plate; The carrier platform is attached to mixture 1, the elution device contains eluent, and the cover plate is attached to mixture 2; The mixture 1 is composed of agarose, hematoxylin solution, and water; The eluent consists of an ethanol solution, eosin staining solution, and water. The mixture 2 is composed of neutral resin and environmentally friendly transparent liquid.

[0006] Preferably, the mass-to-volume ratio of agarose, hematoxylin solution and water in mixture 1 is 0.1~1g:1mL:3~5mL; The volume ratio of neutral resin to environmentally friendly transparent liquid in mixture 2 is 3~5:1; The volume ratio of ethanol solution, eosin staining solution and water in the eluent is 2~3:1:1~2.

[0007] Preferably, the concentration of the ethanol solution is >90%.

[0008] Preferably, the support stage is a glass slide, and the support stage is prepared by mixing hematoxylin solution with water, adding agarose, heating to fully dissolve the agarose, and cooling to a semi-gelatinous state to obtain mixture 1. Mixture 1 is then applied to a glass slide to obtain the support stage; the thickness of mixture 1 applied to the glass slide is 1~3mm.

[0009] Preferably, the elution device is a dropper, and the elution device is prepared by mixing ethanol solution, eosin staining solution and water, and then placing the mixture into a dropper to obtain the elution device.

[0010] Preferably, the cover plate is a cover glass slide, and the cover plate is prepared by mixing neutral resin with environmentally friendly transparent liquid to obtain mixture 2, and applying mixture 2 onto the cover glass slide to obtain the cover plate; the thickness of mixture 2 applied to the cover plate is 0.5~2mm.

[0011] Preferably, a thin film is covered on the cover plate of the coating mixture 2.

[0012] This invention provides a method for using the portable cervical exfoliated cell staining kit, comprising the following steps: (1) Apply the sample directly to the support platform, heat it, and after the mixture 1 on the support platform melts, drain it to obtain a support platform with the sample. (2) Use an elution device to clean the support platform with the sample to obtain the cleaned support platform; (3) Place the cover plate on the support platform and let it stand. Then you can use a microscope to observe the sample.

[0013] Preferably, the settling time is 50-70 seconds.

[0014] Compared with the prior art, the present invention has the following beneficial effects: (1) This invention provides a portable cervical exfoliated cell staining kit and its usage method. By making the cervical exfoliated cell staining method portable and providing a corresponding specific staining process, this invention enables timely cervical exfoliated cell staining during gynecological examinations, creating conditions for microscopic diagnosis by pathologists. It mainly solves the shortcomings of current cervical exfoliated cell staining methods, which occupy a large space and are inconvenient to carry, and shortens the time required to obtain the final examination results, thereby enabling patients with positive results to receive timely clinical treatment.

[0015] (2) The present invention uses low-melting-point agarose to make low-melting-point agar gel, which solidifies into a gel at about 30°C and has a melting point of about 65°C. After melting, it can remain liquid for several hours at 37°C. When a 2mm low-melting-point agar gel is applied to a blank glass slide and heated under the slide, the low-melting-point agar gel softens and liquefies at about 60°C. Agarose itself has a certain degree of water absorption and can replace the dehydration effect of ethanol in conventional staining. At the same time, when the mixture liquefies, the hematoxylin solution in it plays a role in staining the nuclei of exfoliated cervical cells. At the same time, both the low-melting-point agar gel and the hematoxylin solution are alkaline substances, and they will not react with each other when placed together, and the chemical properties of the mixture are stable.

[0016] (3) In this invention, a disposable plastic dropper containing 99% ethanol solution is used to further dehydrate the cervical exfoliated cells on the slide, while the commercial eosin staining solution it contains stains the cytoplasm of the cervical exfoliated cells. 99% ethanol solution is a neutral organic solvent, and commercial eosin staining solution is an acidic substance, both of which dissolve well in the ethanol solution. Simultaneously, when the dropper is applied twice to the slide, it effectively eliminates any residual alkaline dye from the previous step. The small amount of residual 99% ethanol solution also promotes the softening of the neutral resin in the next mounting step, enhancing the fusion of the slide and coverslip.

[0017] (4) The neutral resin in this invention serves to fuse the slide and coverslip, while the environmentally friendly clearing solution provides transparency and improves the refractive index, making the exfoliated cervical cells clearer and easier to observe under the microscope. The environmentally friendly clearing solution is volatile; allowing it to stand for 60 seconds after staining effectively promotes its evaporation. Attached Figure Description

[0018] Figure 1 This is a routine staining treatment; Figure 2 Processed by a fully automatic dyeing machine; Figure 3 This is a schematic diagram of the portable cervical exfoliated cell staining kit of the present invention; Figure 4Staining of cervical exfoliated cells using a portable staining device; Figure 5 This is the current standard staining method for cervical exfoliated cells. Detailed Implementation

[0019] This invention provides a portable cervical exfoliated cell staining kit, which includes a support platform, an elution device, and a cover plate; The carrier platform is attached to mixture 1, the elution device contains eluent, and the cover plate is attached to mixture 2; The mixture 1 is composed of agarose, hematoxylin solution, and water; the agarose is preferably low-melting-point agarose. The eluent is composed of an ethanol solution, an eosin staining solution, and water, and the eosin staining solution is preferably a commercially available eosin staining solution. The mixture 2 is composed of neutral resin and environmentally friendly transparent liquid.

[0020] In this invention, the mass-to-volume ratio of agarose, hematoxylin solution and water in mixture 1 is 0.1~1g:1mL:3~5mL, preferably 0.3~0.7g:1mL:2.5~4.5mL, and more preferably 0.5g:1mL:4mL; The volume ratio of neutral resin to environmentally friendly transparent liquid in the mixture 2 is 3~5:1, preferably 3.5~4.5:1, and more preferably 4:1; The volume ratio of ethanol solution, eosin staining solution and water in the eluent is 2~3:1:1~2, preferably 2.2~2.8:1:1.2~1.7, and more preferably 2.5:1:1.5; the concentration of the ethanol solution is >90%, preferably 95%~100%, and more preferably 99%.

[0021] In this invention, the support stage is a glass slide. The preparation method of the support stage is as follows: hematoxylin solution is mixed with water, agarose is added, and heated until the agarose is fully dissolved. After cooling to a semi-gelatinous state, mixture 1 is obtained. Mixture 1 is then applied to a glass slide to obtain the support stage. The heating method is as follows: heat the mixture on high in a microwave oven for 1 minute, stir, then heat on high in the microwave oven for 30 seconds, stir again, and then heat on high in the microwave oven for 15 seconds until the agarose is completely dissolved. If it is not dissolved, the heating method is repeated 3 times until the agarose is completely dissolved. The thickness of mixture 1 applied to the glass slide is 1~3 mm, preferably 1.5~2.5 mm, and more preferably 2 mm.

[0022] In this invention, the elution device is a dropper, and the elution device is prepared by mixing an ethanol solution, an eosin staining solution and water, and then placing the mixture into a dropper to obtain the elution device; the dropper is preferably a disposable plastic dropper.

[0023] In this invention, the cover plate is a cover glass slide, and the cover plate is prepared by mixing neutral resin with environmentally friendly transparent liquid to obtain mixture 2, and applying mixture 2 onto the cover glass slide to obtain the cover plate; the thickness of mixture 2 applied to the cover plate is 0.5~2mm, preferably 0.7~1.5mm, more preferably 1mm, and preferably a thin film is covered on the cover plate with mixture 2 applied, the thin film being preferably a transparent PE plastic film.

[0024] This invention provides a method for using the portable cervical exfoliated cell staining kit, comprising the following steps: (1) Apply the sample directly to the support platform, heat it, and after the mixture 1 on the support platform melts, drain it to obtain a support platform with the sample. (2) Use an elution device to clean the support platform with the sample to obtain the cleaned support platform; (3) Place the cover plate on the support platform and let it stand. Then you can use a microscope to observe the sample.

[0025] In this invention, the heating method is as follows: the support platform is placed at a distance of 18-22cm from the electric wire furnace, preferably 19-21cm, and more preferably 20cm.

[0026] In this invention, the cleaning method is as follows: the eluent from the elution device is dropped onto the support stage containing the sample. During the first drop, the mixture 1 is washed away. During the second drop, the glass slide is placed flat, and the eluent is kept on the support stage for 30 seconds. The eluent is then poured off to complete the cleaning.

[0027] In this invention, the step of placing the cover plate on the support platform preferably involves removing the film covering the cover plate before placing the cover plate on the support platform; the settling time is 50-70 seconds, preferably 55-65 seconds, and more preferably 60 seconds.

[0028] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0029] Example: Preparation of a portable cervical exfoliated cell staining kit

[0030] Preparation of the stage (A): A 2mm layer of mixture 1 was uniformly coated on a blank glass slide (produced by Wuhan Bominghui Biotechnology Co., Ltd., Bioland frosted glass slide, size 75×25×1mm, model GS7105-7525).

[0031] The preparation process and proportions of Mixture 1 are as follows: Add 20 mL of hematoxylin solution (produced by Nanjing Senbeijia Biotechnology Co., Ltd., Ehrlich hematoxylin staining solution, model BP-DL003) to a 250 mL Erlenmeyer flask, then add 80 mL of distilled water, and then slowly add 10 g of low-melting-point agarose (produced by Hubei Huiheyuan Chemical Co., Ltd., model agarose ME). The low-melting-point agarose should be added while stirring (1 s / cycle, stirring for 30 s) to prevent aggregation. After thorough stirring, place the Erlenmeyer flask in a microwave oven (Galanz microwave oven, model P20D20TL-D4) and heat on high for 1 minute, stirring thoroughly. Then heat on high for another 30 seconds, stirring thoroughly again, and then heat on high for another 15 seconds. Continue until the low-melting-point agarose is fully dissolved. If it is not fully dissolved and solid particles are still visible, repeat the above heating process three times. After the low-melting-point agarose is fully dissolved, the mixture is allowed to cool naturally to about 65°C and become semi-gelatinous. It is then evenly spread on one side of a blank glass slide to a thickness of 2 mm. After spreading, it is allowed to cool naturally to room temperature for later use.

[0032] Preparation of the elution device (B): 99% ethanol solution (10L), commercial eosin staining solution (CD-RS-011), and distilled water were placed in a disposable plastic dropper (produced by Taizhou Yuze Medical Supplies Co., Ltd., specification 10mL).

[0033] The preparation process and ratio of the eluent are as follows: Take 20 mL of commercial eosin staining solution and add it dropwise to 50 mL of 99% ethanol solution while stirring (1 s / cycle, stirring for 30 s). After stirring thoroughly, add 30 mL of distilled water and stir thoroughly. Then, take 20 mL of the solution and place it in a dropper-type plastic vial for later use.

[0034] Preparation of the cover plate (C): Apply a layer of mixture 2 about 1mm thick evenly on the empty cover glass slide (produced by Guangzhou Dianrui Chemical Glass Experimental Instrument Co., Ltd., specifications 24×50×0.15mm, model 010408001), and then attach a layer of transparent PE plastic film (produced by Dongguan Xincan Packaging Products Co., Ltd., specifications 30mm wide, thickness 2 mil).

[0035] The preparation process and proportion of mixture 2 are as follows: Take 40mL of neutral resin (produced by Nanchang Yulu Experimental Equipment Co., Ltd., batch number 200801), and add 10mL of environmentally friendly transparent liquid (produced by Guangzhou Cida Biotechnology Co., Ltd., model Bio-CD2339) evenly to it, stirring thoroughly while adding the liquid.

[0036] To obtain a portable cervical exfoliated cell staining kit, see [link / reference]. Figure 3 .

[0037] Experimental Example

[0038] Using the portable cervical exfoliated cell staining kit obtained in the examples, cervical exfoliated cells from 50 subjects who underwent outpatient physical examinations at the Affiliated Hospital of Yangzhou University from May to June 2025 were stained using conventional staining methods. Figure 1 As a control, the results are shown in [reference needed]. Figure 4 and Figure 5 .

[0039] Instructions for use of the portable cervical exfoliative cell staining kit: (1) When a gynecologist performs a cervical examination on a patient, a smear is taken at the target area of ​​the cervix. The examination end of the smear is then directly applied back and forth three times on the surface of the substrate to which mixture 1 is applied. The smeared slide is then heated on a flameless electric furnace, ensuring that the smeared side is facing up and that the slide is about 20 cm above the furnace. The slide is heated until the adhesive on it melts. After the adhesive melts, the heating is maintained for about 90 seconds. The melted adhesive is then drained off and set aside.

[0040] (2) After the melted adhesive is drained off, the eluent is dropped onto the glass slide using an elution device. The residual adhesive is washed off during the first drop. During the second drop, the glass slide is laid flat so that the eluent remains on the glass slide for about 30 seconds. Then the eluent is drained off.

[0041] (3) Remove the cover plate, peel off the transparent PE plastic film on the surface, align it with the target area on the slide, and place the cover plate on top. At the same time, observe the upper and lower edges of the cover plate to ensure that it does not exceed the corresponding upper and lower edges of the slide. Gently press the cover plate to squeeze out the small air bubbles under the cover plate. After standing for 60 seconds, it can be observed under a microscope.

[0042] The portable cervical exfoliated cell staining kit of this invention was used to stain cervical exfoliated cells from 50 subjects. The results showed that all 50 cervical exfoliated cells were satisfactorily stained under the microscope. The nuclei were blue, the cytoplasm was red, the contrast between the nuclei and cytoplasm was clear, the cell outlines were clear, and the cell membranes were visible, meeting the requirements for microscopic diagnostic analysis.

[0043] Depend on Figure 4 and Figure 5 The comparison shows that the staining effect of the cervical exfoliated cells after staining with the portable cervical exfoliated cell staining kit is satisfactory, achieving the staining effect of the current routine staining method for cervical exfoliated cells.

[0044] The portable cervical exfoliative cell staining kit of the present invention has the advantages of being space-saving and easy to carry, and the consumables are inexpensive. When manufactured in large quantities, the cost of consumables per kit is approximately RMB 1.5, making it suitable for widespread use in rural and remote areas and meeting the requirements for extensive and regular cervical cancer screening in my country.

[0045] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A portable kit for staining cervical exfoliated cells, characterized in that, It comprises a bearing platform, an elution device and a cover plate. The bearing platform is attached with mixture 1, the elution device contains eluent, and the cover plate is attached with mixture 2. The mixture 1 is composed of agarose, hematoxylin solution and water. The eluent is composed of ethanol solution, eosin staining solution and water. The mixture 2 is composed of neutral gum and environment-friendly transparent liquid.

2. The portable cervical exfoliated cell staining kit of claim 1, wherein, The mass-volume ratio of agarose, hematoxylin solution and water in the mixture 1 is 0.1-1g:1mL:3-5mL. The volume ratio of neutral gum and environment-friendly transparent liquid in the mixture 2 is 3-5:

1. The volume ratio of ethanol solution, eosin staining solution and water in the eluent is 2-3:1:1-2.

3. The portable cervical exfoliated cell staining kit according to claim 1 or 2, wherein, The concentration of the ethanol solution is >90%.

4. The portable cervical exfoliated cell staining kit of claim 1, wherein, The bearing platform is a glass slide, and the preparation method of the bearing platform is as follows: hematoxylin solution and water are mixed, then agarose is added and heated to fully dissolve the agarose, and then the mixture 1 is obtained by cooling to semi-gel state, and the mixture 1 is applied to the glass slide to obtain the bearing platform. The thickness of the mixture 1 applied to the glass slide is 1-3mm.

5. The portable cervical exfoliated cell staining kit of claim 1, wherein, The elution device is a dropper, and the preparation method of the elution device is as follows: ethanol solution, eosin staining solution and water are mixed, and then the mixture is filled into the dropper to obtain the elution device.

6. The portable cervical exfoliated cell staining kit of claim 1, wherein, The cover plate is a cover glass, and the preparation method of the cover plate is as follows: neutral gum and environment-friendly transparent liquid are mixed to obtain mixture 2, and then the mixture 2 is applied to the cover glass to obtain the cover plate. The thickness of the mixture 2 applied to the cover plate is 0.5-2mm.

7. The portable cervical exfoliated cell staining kit of claim 6, wherein, A layer of film is covered on the cover plate with the mixture 2 applied thereon.

8. The method of using a portable cervical exfoliated cell staining kit according to any one of claims 1 to 7, wherein, It comprises the following steps: (1) The sample is directly applied to the bearing platform, heated, and then the mixture 1 on the bearing platform is drained after melting to obtain the bearing platform with the sample; (2) The bearing platform with the sample is cleaned using the elution device to obtain the cleaned bearing platform; (3) The cover plate is covered on the bearing platform, and then the sample is observed using a microscope.

9. The method of use of claim 8, wherein, The standing time is 50-70s.

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