A gram staining cartridge and method of use thereof

Abstract

The application discloses a gram staining box, which is characterized by four independent containing cavities for preloading corresponding dyeing liquid separated by a partition plate in the box body, a soft film with a partitioned tearable sealing film and a corresponding jack for each containing cavity, and is integrated into a one-time staining box. The slide is inserted into each containing cavity through the jack to complete step-by-step staining. The gram staining box integrates four kinds of dyeing liquids required for gram staining into a single box body, stores the dyeing liquids in a closed mode, and is double-sealed by the soft film and the partitioned tearable sealing film. Only a single and corresponding jack is opened step by step for the staining operation, the area of the dyeing liquid exposed to the air is greatly reduced, the leakage of volatile and irritating gas of the dyeing liquid is inhibited from the source, harmful gas is prevented from accumulating in the closed microbiological laboratory, the occupational injury of the respiratory tract and the body of the experimental personnel caused by the chemical volatile substances is reduced, the experimental operation environment is optimized, and the operation safety is improved.

Description

Technical Field

[0001] This invention relates to the field of diagnostic reagent kit technology, and in particular to a Gram staining kit and its method of use. Background Technology

[0002] Gram staining is a fundamental test for bacterial typing and identification in microbiology laboratories. Currently, the standard laboratory procedure is in-situ staining on a glass slide. After fixing the specimen smear, the operator sequentially opens the separately dispensed primary stain, mordant, destaining solution, and counterstain, pouring each stain directly onto the bacterial film surface of the slide. A rubber bulb is used to smooth the stain, and after standing for the prescribed time, the slide is rinsed with running water. This four-step staining process is repeated. All four types of stains used in Gram staining are volatile, and the volatilized chemical components produce a pungent, irritating odor. To ensure specimen testing is not affected by external bacteria, microbiology laboratories cannot achieve full-area ventilation. The volatile gases from the stains accumulate in the experimental space, causing laboratory air pollution. Long-term inhalation of these volatile agents can irritate the respiratory tract of operators, damage their health, and pose a high occupational health risk. Meanwhile, the operation mode of opening the bottle cap and pouring the dye solution will continuously increase the amount of dye solution volatilization. The use of open reagent bottle mouth will also cause the dye solution to be contaminated and deteriorated by environmental bacteria, affecting the dyeing accuracy. After each process of washing the slides, excess dye solution will be attached to the surface. During the transportation process, residual liquid dripping will contaminate the laboratory table surface. Moreover, the residual dye solution carried by the slides can easily cause cross-color mixing between processes, further reducing the dyeing qualification rate.

[0003] It is evident that existing technologies still need improvement and enhancement. Summary of the Invention

[0004] In view of the shortcomings of the prior art, the purpose of this invention is to provide a Gram staining kit, which aims to solve the technical problems of traditional Gram staining reagents being stored in separate bottles and subjected to multiple staining sessions, which can easily lead to the volatilization of the dye solution, contaminating the experimental space and endangering the health of personnel.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: A Gram staining box includes a transparent box body and a box cover that can be detachably attached to the box body. The box body has three partitions arranged parallel to each other inside. The edges of each partition are sealed to the inner wall of the box body. The three partitions divide the internal space of the box body into four independent receiving cavities. The four receiving cavities are respectively used to hold the primary staining solution, mordant solution, decolorizing solution and counterstaining solution required for Gram staining. A soft membrane is fixedly installed at the open end of the box body. The soft membrane seals and covers the opening of the box body and is located between the box body and the lid. Four insertion holes are opened through the soft membrane. The four insertion holes are respectively set to correspond one-to-one with the four receiving cavities. The insertion holes are used for the glass slides to be stained to pass through and extend into the corresponding receiving cavities.

[0006] The Gram staining box is described in which the inner wall contour of the insertion hole of the soft membrane is adapted to the cross-sectional contour of the glass slide. The inner diameter of the insertion hole is larger than the cross-sectional size of the glass slide and smaller than the opening size of the corresponding receiving cavity, so that the glass slide can be inserted and limited and the upper opening of the receiving cavity is blocked.

[0007] The Gram staining box further includes a peelable sealing film, which covers the side of the soft film away from the box body. The four edges of the sealing film are fixedly connected to the four edges of the soft film, and the sealing film is used to cover the various holes of the soft film.

[0008] The Gram staining box has three connection points on the side of the sealing film facing the soft film. The three connection points are arranged one-to-one in the gap between two adjacent insertion holes, so that the sealing film can be partitioned and peeled off.

[0009] The Gram staining box has a lug extending from one edge of the sealing film. The lug is located on the side of the receiving cavity corresponding to the primary staining solution and is used to open the box by force when the sealing film is peeled off.

[0010] The Gram staining box has four markings, A, B, C, and D, on its outer wall. Mark A corresponds to the primary staining solution chamber, mark B corresponds to the mordant staining solution chamber, mark C corresponds to the decolorizing solution chamber, and mark D corresponds to the secondary staining solution chamber.

[0011] The Gram staining box has a snap-fit ​​ring protruding from the outer wall of the box body, and the snap-fit ​​ring is located near the edge of the box body opening.

[0012] The Gram staining box has a damping ring protruding inward from the inner side wall of the lid. The damping ring is located near the lower edge of the lid. The damping ring and the snap-fit ​​ring are detachably and sealingly snapped together, so as to realize the detachable snap-fit ​​between the lid and the box body.

[0013] A method for using a Gram staining box, based on the Gram staining box described above, includes the following steps: Open the box lid, peel off the sealing film in sections, exposing only the insertion hole corresponding to the primary staining solution container cavity. Insert the prepared and fixed glass slide to be stained into the container cavity through the insertion hole, so that the bacterial film area of ​​the glass slide is soaked in the primary staining solution to complete the primary staining. Take out the glass slide and wash it with water. Continue peeling off the sealing film in sections to expose the insertion holes of the corresponding mordant solution receiving chambers. Insert the washed glass slide into the corresponding receiving chamber to immerse it in the mordant solution and complete the mordant treatment. Remove the glass slide and wash it with water. Continue peeling off the sealing film in sections to expose the insertion holes of the corresponding decolorizing liquid receiving chambers. Insert the washed glass slides into the corresponding receiving chambers to immerse them in the decolorizing liquid and complete the decolorization. Remove the glass slides and wash them with water. Completely peel off the sealing film to expose the insertion hole of the corresponding counterstaining solution cavity. Insert the washed glass slide into the corresponding cavity to immerse it in the counterstaining solution and complete the counterstaining process. Remove the stained slides and wash them with water. Reattach the sealing film to the soft film surface, seal the lid and body of the box, and finally dispose of them in the designated waste collection container for centralized disposal.

[0014] The Gram staining box described herein is used in a way that, when the slide is removed after each staining step, the gap between the wall of the corresponding insertion hole and the outer wall of the slide can be used to scrape off and retain excess staining solution adhering to the surface of the slide, thus avoiding cross-contamination between different staining solutions.

[0015] Beneficial effects: This invention provides a Gram staining box, which is divided into four independent chambers pre-filled with corresponding staining solutions by a partition plate inside the box. It is combined with a soft film with a tearable sealing film and a socket corresponding to each chamber to form an integrated disposable staining box. Slides are inserted into each chamber in sequence through the sockets to complete the step-by-step staining. This Gram staining box integrates the four types of staining solutions required for Gram staining into a single pre-filled box. The staining solutions are stored in a completely sealed environment, with double-layer sealing protection from a soft membrane and a removable, partitioned sealing film. Staining operations are performed only by opening a single alignment port in each step, significantly reducing the area of ​​the staining solution exposed to air. This inhibits the leakage of volatile and irritating gases from the staining solution from the source, preventing the accumulation of harmful chemical odors in the closed microbiology laboratory. It also reduces occupational hazards to the respiratory tract and body of laboratory personnel caused by volatile chemical substances, optimizes the experimental operating environment, and improves operational safety. At the same time, the sealing film can be opened and closed in sections to match the predetermined step-by-step operation sequence of Gram staining. The unused staining solution chamber remains completely sealed, preventing the staining solution from being contaminated and deteriorated by environmental bacteria, ensuring the accuracy of staining results, and further reducing the loss of idle staining solution due to evaporation. During slide insertion and removal, the wall of the insertion hole can scrape off excess dye from the slide surface, preventing residual liquid from dripping onto the work surface and cross-contamination between procedures, reducing reagent contamination on the work surface, and eliminating the need for operators to repeatedly pour and drip dye. The integrated structure eliminates the need for multiple sets of independent reagent bottles, reducing the space occupied by experimental consumables. The sealed snap-fit ​​structure between the box and the lid provides better sealing during storage and transportation. In addition, the product is designed for single use, which completely avoids the problem of dye reuse and deterioration, thus improving the standardization, convenience, and personnel protection of Gram staining operations. Attached Figure Description

[0016] Figure 1 This is an exploded structural diagram of the Gram staining box provided by the present invention; Figure 2 A cross-sectional view of the Gram staining box provided by the present invention; Figure 3 Provided by the present invention Figure 2 A schematic diagram of the partial structure of A in the middle.

[0017] Figure label: 1—Box body 2—Box lid 3—Soft film 4—Sealing membrane; 11—Separator; 12—Receiving cavity 13—Snap-fit ​​ring; 21—Damping ring; 31—Socket 41—Connector position; 42—Lug. Detailed Implementation

[0018] This invention provides a Gram staining kit and its method of use. To make the objectives, technical solutions, and effects of this invention clearer and more explicit, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only for explaining the invention and are not intended to limit the invention.

[0019] In the description of this invention, it should be understood that the terms "upper," "lower," "left," and "right," etc., indicating orientation or positional relationships based on the orientation or positional relationships shown in the accompanying drawings, are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or a specific orientational structure and operation. Therefore, they should not be construed as limitations on the invention. Furthermore, "first" and "second" are only for descriptive purposes and should not be construed as indicating or implying relative importance or implicitly specifying the number of indicated technical features. Therefore, a feature defined with "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of this invention, unless otherwise stated, "multiple" means two or more.

[0020] Please see Figures 1 to 2 As shown, the present invention provides a Gram staining box, including a box body 1 made of transparent material and a matching box lid 2. The box lid 2 and the opening of the box body 1 are assembled by a detachable snap-fit ​​structure. The box body 1 is made of rigid transparent plastic and is injection molded in one piece, which makes it easy for operators to directly observe the remaining amount of dye in each containing cavity 12 and the immersion status of the glass slide from the outside of the box body 1. The box lid 2 is made of the same plastic material and is fastened to the outside of the box body 1 during the product storage and transportation stage to protect the overall structure and prevent the box body 1 from being damaged and leaking due to external impact. The box body 1 has three parallel partition plates 11 arranged inside. The partition plates 11 are injection molded into an integral structure with the inner wall of the box body 1. The edges of each partition plate 11 are seamlessly sealed to the inner wall of the box body 1. The three partition plates 11 divide the internal space of the box body 1 into four independent and isolated cavities 12. The four cavities 12 are arranged in sequence along the width of the box body 1. During the product packaging stage, they are respectively filled with the primary dyeing solution, mordant solution, decolorizing solution and re-dyeing solution required for Gram staining. The dyeing solutions inside each cavity 12 are completely separated by the partition plates 11, so different dyeing solutions will not mix during storage and transportation.

[0021] An elastic soft membrane 3 is adhered and fixed to the open end face of the box body 1. The soft membrane 3 is made of silicone film that is resistant to corrosion by various dyeing reagents. The soft membrane 3 covers and seals the entire open end face of the box body 1. The soft membrane 3 is located between the box body 1 and the box cover 2. It relies on its own sealing property to prevent the dye solution in the receiving cavity 12 from evaporating and leaking out from the opening position. Four insertion holes 31 are opened through the soft membrane 3. The four insertion holes 31 are respectively set vertically and vertically to the four receiving cavities 12. The insertion holes 31 are used for the glass slide to be stained to pass through and extend into the corresponding receiving cavity 12. The inner contour of the insertion hole 31 of the soft membrane 3 is adapted to the cross-sectional contour of a conventional pathological slide. The inner diameter of the insertion hole 31 is slightly larger than the cross-sectional size of the slide and smaller than the opening size of the corresponding receiving cavity 12. This ensures that the slide can be smoothly inserted vertically into the insertion hole 31 and sink to the bottom of the staining solution in the receiving cavity 12, so that the bacterial film coated on the slide is fully immersed in the staining solution to complete the staining reaction. At the same time, when the slide is pulled out upwards, the inner wall of the insertion hole 31 can be used to scrape and retain the excess staining solution adhering to the outer wall of the slide, allowing the scraped-off staining solution to fall back into the original receiving cavity 12.

[0022] In actual use, first remove the outer box cover 2, then tear open the matching partition sealing film 4 above the soft film 3 in sequence, and expose the corresponding insertion holes 31 according to the staining sequence of primary staining, mordant staining, decolorization and restaining. Insert the fixed specimen slides into each receiving cavity 12 in sequence to complete the distribution staining. After the entire staining operation is completed, the whole box of instruments is disposed of as a disposable medical consumable.

[0023] Please see Figures 1 to 3 As shown, a peelable sealing film 4 is attached to the surface of the soft film 3. The sealing film 4 covers the side of the soft film 3 away from the box body 1. The sealing film 4 covers all the areas where the sockets 31 are located. The overall size of the sealing film 4 is consistent with the outer contour of the soft film 3. Its four edges are glued and fixed around the four sides of the soft film 3. The sealing film 4 and the soft film 3 are reliably connected by the glued edges. Under normal conditions, the sealing film 4 is completely attached to the surface of the soft film 3 and fully covers all the sockets 31, sealing all the sockets 31. This can isolate the outside air and dust during the product storage and transportation stage, prevent the various dyes pre-filled in the containment cavity 12 from evaporating and being lost through the sockets 31, and prevent external pollutants from entering the cavity through the sockets 31 and causing the dyes to deteriorate and fail.

[0024] In addition to the perimeter sealing and bonding, the sealing film 4 and the soft film 3 are connected by three connection positions 41 on the side of the sealing film 4 facing the soft film 3. The three connection positions 41 are set one by one in the gap between two adjacent insertion holes 31. The remaining area of ​​the film body and the soft film 3 are separable. When using it, the operator can tear off the sealing film 4 in sections according to the dyeing sequence. Each section torn off will expose a single insertion hole 31. The untorn part will still keep the remaining insertion holes 31 sealed and will not expose the insertion holes 31 corresponding to the subsequent dyeing solutions in advance. This ensures that the four dyeing solutions are always in a sealed and preserved state before the corresponding insertion holes 31 are used. After all dyeing processes are completed, the sealing film 4 does not need to be restored and pasted. The dyeing box after dyeing is discarded along with the consumables.

[0025] The sealing film 4 has an integrally formed lug 42 extending outward from one of its edges along its length. The lug 42 is located near the receiving cavity 12 corresponding to the primary dyeing solution and at a corner. The lug 42 and the sealing film 4 are made of the same thin film material and are cut together. This lug extends outward from the outer contour of the soft film 3 without being covered by the soft film 3. In normal storage, the lug 42 naturally lies flat against the outer edge of the box 1 and does not occupy the storage space of the box 1. When the operator needs to peel off the sealing film 4, they can directly pinch the exposed lug 42 and pull it upward. Using the force point formed by the lug 42, the end of the sealing film 4 can be easily pulled up. The film can be torn apart in sections along the separation position between the adjacent insertion holes 31, gradually revealing the insertion holes 31 corresponding to primary dyeing, mordant dyeing, decolorization, and re-dyeing. The sealing film 4 can be peeled off in sections without the need for auxiliary tools such as blades and tweezers, simplifying the operation steps of opening the lid and removing the film.

[0026] Please see Figure 1 As shown, the outer wall of box 1 is marked with four labels: A, B, C, and D. Label A corresponds to the primary dyeing solution chamber 12, label B corresponds to the mordant dyeing solution chamber 12, label C corresponds to the decolorizing solution chamber 12, and label D corresponds to the secondary dyeing solution chamber 12. Each label is formed on the corresponding outer position of the transparent outer wall of box 1 by injection molding or ink printing. The labels correspond one-to-one with the inner cavities. Operators can simultaneously compare the labels with the positions of the dyeing solutions inside the box 1 through the transparent shell from the outside. Combined with the segmented tearing sequence of the sealing film 4, the type of dyeing solution corresponding to each insertion hole 31 can be quickly distinguished, avoiding the problem of mistakenly using the wrong insertion hole 31 or immersing the glass slide into the wrong dyeing solution during use, and further standardizing the dyeing operation sequence.

[0027] Please see Figure 3As shown, a snap-fit ​​protrusion 13 is integrally injection-molded around the outer wall of the box body 1 near the upper opening. The snap-fit ​​protrusion 13 is continuously arranged along the outer perimeter of the box body 1, and its cross-section has an outwardly convex arc structure. The entire ring is located below the position where the soft membrane 3 is installed. The matching box cover 2 is an inverted cover structure. A damping protrusion 21 is integrally formed inwardly near the lower opening on the inner wall of the box cover 2. The damping protrusion 21 is wrapped around the entire inner side of the box cover 2, and the damping protrusion 21 adopts a slightly interference fit design. During assembly and storage, the lid 2 is fastened from top to bottom onto the outside of the opening of the box body 1. The damping ring 21 slides down the outer wall of the box body 1 and elastically clamps onto the lower side of the locking ring 13. The sealing is achieved by the clamping force generated by the plastic deformation of the damping ring 21 itself, thus firmly limiting the lid 2 to the outside of the box body 1. The lid 2 covers the outside of the soft film 3 and the sealing film 4, which can protect the film from being scratched and damaged during storage and transportation, effectively avoiding the failure of the seal and leakage of dye due to impact. When the operator performs the dyeing operation, the lid 2 can be lifted upward to overcome the damping resistance and remove the lid 2. The lid 2 is placed separately throughout the dyeing process. Since the product is a disposable item, the lid 2 is disposed of as medical waste along with the whole box of dyeing boxes after use.

[0028] A method for using a Gram staining box, based on the Gram staining box described above, includes the following steps: When using a pre-filled disposable Gram staining box, first hold the side wall of the box body 1 and pull the lid 2 upwards. The damping protrusion 21 on the inner side of the lid 2 will be stretched open by the external force and deformed, disengaging from the snap-fit ​​protrusion 13 on the outer side of the box body 1, thus successfully releasing the snap-fit ​​protection between the lid 2 and the box body 1, and fully exposing the sealing film 4 attached to the outer side of the end face of the soft film 3. The operator pinches the lug 42 of the sealing film 4 on the side of the primary staining solution and pulls it upwards. The sealing film 4 is peeled off in sections by relying on the connection points between the adjacent insertion holes 31. The first peeling only lifts the first section of the film, exposing only the primary staining solution insertion hole 31 corresponding to the letter A. The remaining section of the sealing film 4 is still tightly attached to the soft film 3, sealing the other three insertion holes 31, effectively preventing the mordant solution, decolorizing solution and restaining solution from evaporating or being contaminated by the environment in the early stage of operation. The operator takes a specimen slide that has been smeared and flame-fixed, keeps the bacterial film side facing the inside of the receiving cavity 12, and slowly lowers it vertically into the exposed insertion hole 31 until the bacterial film on the slide is completely submerged below the surface of the primary staining solution. The slide is then left to stand and soak for the prescribed duration according to Gram staining standards to complete the primary staining process. During the process of lifting the slide upwards, the gap between the inner wall of the insertion hole 31 and the side wall of the slide is appropriate, and the edge of the insertion hole 31 can scrape off the excess primary staining solution trapped on the outer wall of the slide. The scraped-off staining solution falls back into the primary staining solution receiving cavity 12 below through the insertion hole 31 without dripping or overflowing. The slide is then moved to a water tank and gently rinsed with running water to remove the free staining solution on the surface of the slide.

[0029] After the initial dyeing and washing process is completed, hold the sealing film 4 again and continue to peel off the sealing film 4 in sections to expose the mordant liquid insertion hole 31 corresponding to the B mark on the side wall of the box body 1. Insert the washed and drained glass slide vertically into the insertion hole 31 and immerse it in the mordant liquid in the cavity. Let it stand to complete the mordant reaction. When the glass slide is pulled out upward, scrape off the excess mordant liquid by relying on the wall of the insertion hole 31. The excess dye liquid flows back into the original cavity and the glass slide is washed again.

[0030] Following the same operational logic, the sealing film 4 is torn open in sections to expose the decolorizing liquid insertion hole 31 corresponding to the C mark. The glass slide is then soaked in the decolorizing liquid, and any residual liquid is scraped off and the slide is washed with water when it is removed.

[0031] After the first three staining steps are completed, completely peel off all the remaining sealing film 4 to expose the counterstaining solution insertion hole 31 corresponding to the D mark. Insert the glass slide that has been decolorized and washed into the cavity to immerse it in the counterstaining solution. When removing the slide, the insertion hole 31 will retain any excess counterstaining solution again.

[0032] After all staining steps are completed, the finished slides are removed and given a final rinse. Following the operating instructions, the remaining sealing film 4 is temporarily applied back onto the surface of the soft film 3. Then, the lid 2 is fastened from top to bottom to the opening of the box body 1, ensuring the inner damping ring 21 of the lid 2 is securely fastened to the locking ring 13 of the box body 1, thus completing the sealing of the box body 1. Because this staining box is a disposable design, the four staining solutions inside cannot be reused after step-by-step staining. The entire box body 1, along with the lid 2, is disposed of in a dedicated medical waste collection container for unified and harmless treatment. Throughout the entire operation, the A, B, C, and D markings on the outside of the box body 1 are used to quickly verify the insertion holes 31 and the type of staining solution, effectively preventing the mixing of staining solutions caused by incorrect slide insertion. The real-time scraping structure of the insertion holes 31 eliminates cross-contamination caused by different staining solutions carried by the slides, simplifying the cumbersome process of repeatedly using multiple reagent bottles in traditional staining methods.

[0033] It is understood that those skilled in the art can make equivalent substitutions or modifications to the technical solution and inventive concept of the present invention, and all such substitutions or modifications should fall within the protection scope of the appended claims.

Claims

1. A Gram staining box, comprising a transparent box body (1) and a box lid (2) detachably attached to the box body (1), characterized in that, The box (1) has three partition plates (11) arranged in parallel intervals inside. The edges of each partition plate (11) are sealed to the inner wall of the box (1). The three partition plates (11) divide the internal space of the box (1) into four independent cavities (12). The four cavities (12) are used to hold the primary staining solution, mordant solution, decolorizing solution and counterstaining solution required for Gram staining, respectively. A soft membrane (3) is fixedly installed at the opening end of the box body (1). The soft membrane (3) seals and covers the opening of the box body (1) and is located between the box body (1) and the box cover (2). Four insertion holes (31) are opened through the soft membrane (3). The four insertion holes (31) are respectively set to correspond one-to-one with the four receiving cavities (12). The insertion holes (31) are used for the glass slide to be stained to pass through and extend into the corresponding receiving cavity (12).

2. The Gram staining kit according to claim 1, characterized in that, The inner wall contour of the insertion hole (31) of the soft membrane (3) is adapted to the cross-sectional contour of the glass slide. The inner diameter of the insertion hole (31) is larger than the cross-sectional size of the glass slide and smaller than the opening size of the corresponding receiving cavity (12), so that the glass slide can be inserted and limited and the upper opening of the receiving cavity (12) is blocked.

3. The Gram staining box according to claim 1, characterized in that, It also includes a peelable sealing film (4), which covers the side of the soft film (3) away from the box body (1). The four edges of the sealing film (4) are fixedly connected to the four edges of the soft film (3). The sealing film (4) is used to cover each of the holes (31) of the sealing soft film (3).

4. The Gram staining box according to claim 3, characterized in that, The sealing film (4) has three connection positions (41) on the side facing the soft film (3). The three connection positions (41) are set in the gap between two adjacent holes (31) in a one-to-one correspondence, so that the sealing film (4) can be partitioned and peeled off in sections.

5. The Gram staining box according to claim 4, characterized in that, One edge of the sealing film (4) extends to form a lug (42). The lug (42) is located on the side of the receiving cavity (12) corresponding to the initial dyeing liquid, and is used to open the sealing film (4) by force when peeling it off.

6. The Gram staining kit according to claim 1, characterized in that, The outer wall of the box (1) is marked with four labels: A, B, C and D. Label A corresponds to the primary dyeing solution container (12), label B corresponds to the mordant solution container (12), label C corresponds to the decolorizing solution container (12), and label D corresponds to the secondary dyeing solution container (12).

7. The Gram staining box according to claim 1, characterized in that, A snap-fit ​​ring (13) is formed on the outer side wall of the box body (1), and the snap-fit ​​ring (13) is set near the edge of the opening of the box body (1).

8. The Gram staining box according to claim 7, characterized in that, A damping protrusion ring (21) is provided on the inner side wall of the lid (2). The damping protrusion ring (21) is located near the lower edge of the lid (2). The damping protrusion ring (21) and the snap-fit ​​protrusion ring (13) are detachably and sealed together, so as to realize the detachable snap-fit ​​between the lid (2) and the box body (1).

9. A method of using a Gram staining box, characterized in that, The Gram staining kit according to any one of claims 1-8 includes the following steps: Open the box lid (2), peel off the sealing film (4) in sections, and expose only the insertion hole (31) of the corresponding primary staining solution container (12). Insert the prepared and fixed glass slide to be stained into the container (12) through the insertion hole (31) so that the bacterial film area of ​​the glass slide is soaked in the primary staining solution to complete the primary staining. Take out the glass slide and wash it with water. Continue to peel off the sealing film (4) in sections to expose the insertion hole (31) of the corresponding mordant solution receiving cavity (12). Insert the water-washed glass slide into the corresponding receiving cavity (12) to soak in the mordant solution and complete the mordant treatment. Take out the glass slide and wash it with water. Continue to peel off the sealing film (4) in sections to expose the insertion hole (31) of the corresponding decolorizing liquid receiving cavity (12). Insert the water-washed glass slide into the corresponding receiving cavity (12) to soak in the decolorizing liquid and complete the decolorization. Take out the glass slide and wash it with water. Completely peel off the sealing film (4) to expose the insertion hole (31) of the corresponding counterstaining solution receiving cavity (12), insert the water-washed glass slide into the corresponding receiving cavity (12) to soak in the counterstaining solution, and complete the counterstaining; Remove the stained slide and wash it with water. Reattach the sealing film (4) to the surface of the soft film (3), seal the lid (2) and the box body (1) together, and finally put it into the designated waste collection container for centralized disposal.

10. The method of using the Gram staining box according to claim 9, characterized in that, When the slide is removed after each staining step, the wall of the corresponding insertion hole (31) is fitted with the outer wall of the slide to scrape off and retain excess staining solution on the surface of the slide, thus avoiding cross-contamination between different staining solutions.

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