A rapid processing method for pathological special staining

Abstract

This invention relates to the fields of pathological techniques and histological staining, specifically a rapid method for special pathological staining. The method involves placing dewaxed and hydrated tissue sections in a KOS multifunctional microwave rapid tissue processor, and then adding reagents for special pathological staining. This invention utilizes microwave energy to enhance the penetration efficiency and reaction rate of staining reagents in tissue, significantly shortening the special staining process while achieving staining quality and tissue morphology effects comparable to traditional staining methods. It is suitable for hospital pathology departments, medical testing institutions, research laboratories, and other locations requiring special pathological staining, and is particularly suitable for applications with high requirements for staining timeliness and batch processing capabilities.

Description

Technical Field

[0001] This invention relates to the fields of pathological techniques and histological staining, specifically, a rapid processing method for special pathological staining. Background Technology

[0002] Specialized pathological staining is an important supplement to routine hematoxylin and eosin (HE) staining. It plays an irreplaceable role in revealing specific chemical components within tissues, extracellular matrix structures, and lesion characteristics, and is widely used in the pathological diagnosis of tumors, fibrosis, metabolic diseases, and myxoid lesions. However, existing specialized staining methods mostly rely on the natural diffusion and reaction processes of reagents under normal pressure, room temperature, or constant temperature conditions. The operation steps are cumbersome, and the staining time for a single slide usually requires 30-65 minutes, which is difficult to meet the increasing workload and turnaround efficiency requirements of current pathology departments.

[0003] In existing technologies, microwave technology has been applied to pathological procedures such as tissue fixation, dehydration, and antigen retrieval, which can shorten processing time to some extent. However, it is mostly limited to a single step or a single application scenario, and there are still problems such as limited reagent penetration efficiency and unstable staining uniformity under normal pressure.

[0004] Therefore, there is still a lack of a technical solution that can significantly shorten the overall process time for various special staining processes while ensuring staining quality and tissue morphology integrity, and that is easy to standardize and promote. Summary of the Invention

[0005] The purpose of this invention is to solve the above-mentioned technical problems and provide a rapid treatment method for special pathological staining (including but not limited to PAS staining, reticular fiber staining, AB staining and Masson trichrome staining) by applying microwave energy to glass slides. By using microwave energy, the penetration efficiency and reaction rate of staining reagents in tissues are improved, and while significantly shortening the special staining treatment time, staining quality and tissue morphology effects comparable to traditional staining methods are obtained.

[0006] To achieve the above objectives, the present invention provides a rapid method for performing special pathological staining using the KOS multifunctional microwave rapid tissue processing instrument, comprising the following steps:

[0007] S1. Tissue sections that have been routinely fixed, embedded in paraffin, and cut, with a section thickness of 3-5 μm;

[0008] S2. Perform routine dewaxing and hydration treatment on the slices;

[0009] S3. Place the dewaxed and hydrated sections in the KOS multifunctional microwave rapid tissue processor and add reagents to the sections for rapid special staining. During the microwave treatment, the temperature is monitored and automatically controlled in real time, and the microwave power is dynamically adjusted according to the set parameters.

[0010] S5. After the staining reaction is completed, perform dehydration, clearing and mounting as needed to obtain stained tissue sections.

[0011] Furthermore, in step S3, the microwave power of the KOS multifunctional microwave rapid tissue processor is 320-400 W. The KOS multifunctional microwave rapid tissue processor can promote rapid and uniform penetration of staining reagents.

[0012] Furthermore, in step S3, the rapid special staining treatment methods include, but are not limited to, PAS staining, reticular fiber staining, AB staining, and Masson tricolor staining.

[0013] Furthermore, in the PAS staining, the periodic acid treatment time is shortened from the conventional 10 min to about 1 min, the Schiff reagent reaction time is shortened from 15 min to about 1 min, and the hematoxylin staining solution reaction time is shortened from 2 min to 20 s.

[0014] In the aforementioned reticular fiber dyeing process, the treatment time for potassium permanganate and silver ammonia solution was reduced from the conventional 5 min to less than 1 min; the treatment time for oxalic acid solution was reduced from 2 min to 10 s; and the treatment time for ferric ammonium sulfate was reduced from 3 min to 20 s.

[0015] In the AB staining process, the staining time for Alsen blue was reduced from the usual 60 min to about 2 min; and the treatment time for nuclear solid red solution was reduced from 5 min to 1 min.

[0016] In the Masson trichrome staining process, the processing time for each major staining step is reduced to 1-2 minutes: the Weigert iron hematoxylin processing time is reduced from the conventional 10 minutes to 2 minutes; the processing time for acid fuchsin, phosphomolybdic acid solution, and aniline blue solution is reduced from the conventional 5 minutes to 1 minute.

[0017] The total processing time for special staining of a single slide using the method of this invention is controlled within 5 minutes.

[0018] This invention utilizes microwave energy to enhance the penetration efficiency and reaction rate of staining reagents in tissues, significantly shortening the time required for specialized staining treatments while achieving staining quality and tissue morphology results comparable to traditional staining methods. It is suitable for hospital pathology departments, medical testing institutions, research laboratories, and other locations requiring specialized pathological staining, and is particularly suitable for applications demanding high staining efficiency and batch processing capabilities. Compared to existing technologies, this invention offers the following advantages:

[0019] 1. Significantly shortens the overall processing time for various special pathological stains, improving the work efficiency of the pathology department;

[0020] 2. While shortening the time, the staining quality, contrast, and specificity are comparable to traditional methods;

[0021] 3. The tissue morphology is well preserved, with no obvious tissue shrinkage, cracks, or section detachment;

[0022] 4. The processing parameters are controllable, with good repeatability and stability, and have good standardization potential;

[0023] 5. It is suitable for various common pathological special staining, has a wide range of applications, and is easy to promote in routine pathology laboratories. Attached Figure Description

[0024] Figure 1 Comparison of processing times between conventional and microwave methods in different special staining processes; This indicates that the difference is statistically significant. P <0.0001).

[0025] Figure 2 A comparison of the effects of conventional methods and microwave treatment methods on different special staining processes. Detailed Implementation

[0026] The specific implementation methods provided by the present invention will be described in detail below with reference to the embodiments.

[0027] Example 1: PAS staining

[0028] S1. Tissue sections that have been routinely fixed, embedded in paraffin, and cut, with a section thickness of 3-5 μm;

[0029] S2. Perform routine dewaxing and hydration treatment on the slices;

[0030] S3. Place the dewaxed and hydrated sections in the KOS multifunctional microwave rapid tissue processor and add reagents for PAS staining. The staining process is monitored and automatically controlled in real time by the system control terminal, with microwave power dynamically adjusted according to the set parameters. The machine power is set to 320-400 W. Record the total processing time from dewaxing to the completion of staining a single section. The processing time and conditions for each staining step are shown in Table 1.

[0031] S5. After the staining reaction is completed, perform dehydration, clearing and mounting as needed to obtain stained tissue sections.

[0032] Example 2: Dyeing of Reticulated Fibers

[0033] S1. Tissue sections that have been routinely fixed, embedded in paraffin, and cut, with a section thickness of 3-5 μm;

[0034] S2. Perform routine dewaxing and hydration treatment on the slices;

[0035] S3. Place the dewaxed and hydrated sections in the KOS multifunctional microwave rapid tissue processor. Add reagents to the sections for reticular fiber staining. The staining process is monitored and automatically controlled in real time by the system control terminal. The microwave power is dynamically adjusted according to the set parameters, and the machine power is set to 320-400 W. Record the total processing time required from dewaxing to the completion of staining a single section. The processing time and conditions for the staining steps are shown in Table 1.

[0036] S5. After the staining reaction is completed, perform dehydration, clearing and mounting as needed to obtain stained tissue sections.

[0037] Example 3: AB staining

[0038] S1. Tissue sections that have been routinely fixed, embedded in paraffin, and cut, with a section thickness of 3-5 μm;

[0039] S2. Perform routine dewaxing and hydration treatment on the slices;

[0040] S3. Place the dewaxed and hydrated sections in the KOS multifunctional microwave rapid tissue processor and add reagents for AB staining. The staining process is monitored and automatically controlled in real time by the system control terminal, and the microwave power is dynamically adjusted according to the set parameters. The machine power is set to 320-400 W. Record the total processing time from dewaxing to the completion of staining a single section. The processing time and conditions for the staining steps are shown in Table 1.

[0041] S5. After the staining reaction is completed, perform dehydration, clearing and mounting as needed to obtain stained tissue sections.

[0042] Example 4: Masson's trichrome staining

[0043] S1. Tissue sections that have been routinely fixed, embedded in paraffin, and cut, with a section thickness of 3-5 μm;

[0044] S2. Perform routine dewaxing and hydration treatment on the slices;

[0045] S3. Place the dewaxed and hydrated sections in the KOS multifunctional microwave rapid tissue processor and add reagents for Masson's trichrome staining. The staining process is monitored and automatically controlled in real time by the system control terminal, with microwave power dynamically adjusted according to set parameters. The machine power is set to 320-400 W. Record the total processing time from dewaxing to the completion of staining a single section. The processing time and conditions for each staining step are shown in Table 1.

[0046] S5. After the staining reaction is completed, perform dehydration, clearing and mounting as needed to obtain stained tissue sections.

[0047] Table 1. Various special staining steps and times

[0048]

[0049] Example 5: Comparison of special staining treatment times using different methods

[0050] Compared with conventional staining methods, microwave treatment significantly reduced the total staining time in PAS staining, reticular fiber staining, AB staining, and Masson's trichrome staining. Figure 1 Conventional methods require approximately 30-65 minutes to complete the aforementioned special staining processes, while microwave treatment can complete them within 10 minutes. Statistical analysis showed that the differences in processing time between the two methods for various special staining processes were statistically significant (P < 0.0001).

[0051] Example 6: Comparison of the quality of special staining methods

[0052] Microscopic observation showed that the tissue structure stained by the microwave rapid processing method was well preserved, with clear cell and tissue morphology, and no obvious tissue shrinkage, cleft formation, or section detachment was observed. Compared with traditional staining methods, no significant differences were found in the localization and morphological appearance of the target structures among various special staining methods. Figure 2 ).

[0053] The preferred embodiments of the present invention have been described in detail above, but the present invention is not limited to the embodiments described. Those skilled in the art can make various equivalent modifications or substitutions without departing from the spirit of the present invention, and these equivalent modifications or substitutions are all included within the scope defined by the claims of this application.

Claims

1. A rapid processing method for special pathological staining, characterized in that, Includes the following steps: S1. Tissue sections that have been routinely fixed, embedded in paraffin, and cut, with a section thickness of 3-5 μm; S2. Perform routine dewaxing and hydration treatment on the slices; S3. Place the dewaxed and hydrated sections in the KOS multifunctional microwave rapid tissue processor and add reagents to the sections for rapid special staining treatment; the microwave power of the KOS multifunctional microwave rapid tissue processor is 320-400 W. S5. After the staining reaction is completed, perform dehydration, clearing and mounting as needed to obtain stained tissue sections.

2. The rapid processing method for special pathological staining according to claim 1, characterized in that, In step S3, rapid special staining methods include, but are not limited to, PAS staining, reticular fiber staining, AB staining, and Masson tricolor staining.

3. The rapid processing method for special pathological staining according to claim 2, characterized in that, In the PAS staining, the periodic acid treatment time was 1 min and the Schiff reagent reaction time was 1 min.

4. The rapid processing method for special pathological staining according to claim 2, characterized in that, In the aforementioned reticular fiber dyeing process, the treatment time for potassium permanganate and silver ammonia solution was 1 min; the treatment time for oxalic acid solution was 10 s; and the treatment time for ferric ammonium sulfate was 20 s.

5. The rapid processing method for special pathological staining according to claim 2, characterized in that, In the AB staining process, the staining time for Elsin Blue was 2 min; the treatment time for Nucleotide Red solution was 1 min.

6. The rapid processing method for special pathological staining according to claim 2, characterized in that, In the Masson trichrome staining process, the treatment time with Weigert iron hematoxylin was 2 min; the treatment time with acid fuchsin, phosphomolybdic acid solution, and aniline blue solution was 1 min.

7. The rapid processing method for special pathological staining according to claim 1, characterized in that, The total processing time for completing the special staining treatment of a single slide should be controlled within 5 minutes.

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