Oral squamous cell carcinoma in-vitro tumor tissue specimen treatment method and application

Abstract

The invention discloses an oral squamous cell carcinoma in-vitro tumor tissue specimen treatment method and application. The oral squamous cell carcinoma in-vitro tumor tissue specimen treatment method specifically comprises the following steps: S1, obtaining an oral squamous cell carcinoma in-vitro tumor tissue specimen; s2, putting the oral squamous cell carcinoma in-vitro tumor tissue specimen into the stationary liquid for fixation; s3, washing with running water 0.5-1 hour before the dehydration procedure is started; s4, fully dehydrating the flushed oral squamous cell carcinoma in-vitro tumor tissue specimen; the invention relates to the technical field of oral squamous cell carcinoma. According to the oral squamous cell carcinoma in-vitro tumor tissue specimen treatment method and application, the low-degree ethanol is used as the start of the first dehydration procedure, and then the concentration of the ethanol is gradually increased, so that the oral squamous cell carcinoma in-vitro tumor tissue specimen is dehydrated more sufficiently, the contractility is small, the tissue texture is not changed, and subsequent slicing is also facilitated; the wax block before slicing is pretreated with an ice-water mixture, so that the hardness of the wax block can be kept proper, and the whole section of the wax block can be uniformly frozen.

Description

Technical Field

[0001] The present invention relates to the technical field of oral squamous cell carcinoma, and in particular to a method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma and its application. Background Art

[0002] Oral squamous cell carcinoma (OSC) is a common malignant tumor of the head and neck. It refers to squamous cell carcinomas that occur in the oral cavity, including the tongue, gums, cheeks, palate, and floor of the mouth. Because the oral mucosa is lined with squamous epithelium, cancers occurring in these areas are called SCCs. SCCs are the most common type of oral cancer. In addition to localized proliferation and erosion of surrounding tissues, causing functional impairment, oral SCCs often also metastasize to the corresponding lymphatic drainage areas of the neck.

[0003] Data show that the five-year survival rate is around 50%, with less than 30% for advanced-stage patients and less than 20% experiencing recurrence. Currently, the treatment for oral squamous cell carcinoma (OSCCC) is a comprehensive sequential approach, primarily surgery, supplemented by chemotherapy and radiotherapy. Studies have shown that local and regional recurrence are a major cause of treatment failure for OSCCC. Complete and thorough resection of the tumor (with negative surgical margins) is key to preventing local recurrence.

[0004] Referring to a Chinese patent, a device for processing in vitro tumor tissue specimens of oral squamous cell carcinoma (Announcement No.: CN113820193A) includes a supporting assembly, a flip assembly, a spray assembly and an immersion box. The supporting assembly is rotatably installed in the immersion box through the flip assembly and the supporting assembly is close to the box opening of the immersion box, and the spray assembly is fixedly installed above the supporting assembly; the supporting assembly includes a cross-clamping rod, a supporting card box and a drive assembly. The supporting card box includes two groups of hollow shells with dry hemispherical shapes, and the two groups of hollow bright bodies are symmetrically arranged on the top of the cross-clamping rod, and there are small holes in a circular equidistant array inside the hollow shell. This device has a simple structure and occupies a small space. It is not only suitable for laboratories, but can also be directly used in clinical practice. It has no limitations and reduces the operator's workload. It supports oral squamous cell carcinoma in vitro specimens at one time without changing their immersion and cleaning positions, reducing the impact of external factors on the in vitro specimens. The operation process is simple and fast, low cost, and does not affect subsequent pathological examinations. However, during the processing of oral squamous cell carcinoma in vitro tumor tissue specimens, it is easy to cause insufficient dehydration, which has a certain impact on the subsequent slicing effect and preservation effect. To this end, the present invention proposes a method and application for processing oral squamous cell carcinoma in vitro tumor tissue specimens to solve the above-mentioned problems. Summary of the Invention

[0005] (1) Technical problems solved

[0006] In view of the deficiencies of the existing technology, the present invention provides a method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma and its application, which solves the problems mentioned in the above background technology.

[0007] (2) Technical solution

[0008] To achieve the above objectives, the present invention is implemented through the following technical solutions: a method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma, specifically comprising the following steps:

[0009] S1. Obtain in vitro tumor tissue specimens of oral squamous cell carcinoma;

[0010] S2. Place the in vitro oral squamous cell carcinoma tumor tissue specimen in a fixative for fixation;

[0011] S3, and rinse with running water 0.5 to 1 hour before the start of the dehydration program;

[0012] S4. Fully dehydrate the washed oral squamous cell carcinoma tumor tissue specimens;

[0013] S5. Finally, the dehydrated oral squamous cell carcinoma in vitro tumor tissue specimen is placed in a solvent for transparency, and then immersed in wax, embedded in paraffin, sliced several times continuously, and baked to obtain the pathological sections.

[0014] Preferably, the oral squamous cell carcinoma in vitro tumor tissue specimen in step S1 includes a primary lesion specimen and a lymph node specimen.

[0015] Preferably, the fixative in step S2 is an ethanol-formaldehyde mixed fixative, wherein the fixation time is 36 to 48 hours.

[0016] Preferably, in the ethanol-formaldehyde mixed fixative, the mass ratio is: 80-90% ethanol, 10-20% formaldehyde.

[0017] Preferably, in step S5, the wax immersion time is 20 to 25 minutes, the wax immersion temperature is 55 to 70° C., and the solvent is a stearic acid paraffin mixture.

[0018] Preferably, the dehydration procedure in step S4 is as follows: running water washing for 0.5 h → 50% ethanol for 1 h → 75% ethanol for 1 h → 85% ethanol for 2 h → 95% ethanol for 2 h → 95% ethanol for 1.5 h → 100% ethanol for 1.5 h.

[0019] Preferably, during paraffin embedding in step S5, the smallest surface of the tissue in contact with the knife is placed at the bottom of the embedding cassette for longitudinal embedding.

[0020] Preferably, in step S5, the wax block is cooled using an ice-water mixture before slicing begins.

[0021] Preferably, the method for processing an in vitro oral squamous cell carcinoma tumor tissue specimen further comprises pretreatment of the tissue specimen: immersing the obtained in vitro oral squamous cell carcinoma tumor tissue specimen in an HCl solution at room temperature for 8 to 25 minutes, and then immersing it in a trypsin-EDTA constant temperature of 33 to 38°C for 15 to 25 minutes.

[0022] The invention relates to an application of an oral squamous cell carcinoma in vitro tumor tissue specimen processed according to the above method in a conventional pathological section examination.

[0023] (3) Beneficial effects

[0024] The present invention provides a method and application for processing ex vivo oral squamous cell carcinoma tumor tissue specimens. Compared with existing technologies, this method and application has the following advantages: By starting with a low-alcohol ethanol dehydration step and then gradually increasing the ethanol concentration, the ex vivo oral squamous cell carcinoma tumor tissue specimen is more fully dehydrated, with minimal shrinkage, preventing changes in tissue texture and facilitating subsequent sectioning. Furthermore, by pre-treating the wax block with an ice-water mixture before sectioning, the wax block maintains an appropriate hardness and evenly freezes the entire cut surface, saving time and effort during sectioning and effectively improving the sectioning effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 A comparison chart of wax blocks pretreated with an improved dehydrated freezing group and an ice-water mixture provided for a method and application of the present invention for processing oral squamous cell carcinoma in vitro tumor tissue specimens. DETAILED DESCRIPTION

[0026] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0027] See also Figure 1 , the embodiment of the present invention provides three technical solutions:

[0028] Example 1:

[0029] A method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma comprises the following steps:

[0030] S1. Obtain in vitro oral squamous cell carcinoma tumor tissue specimens (primary lesion specimens and lymph node specimens);

[0031] S2. Place the in vitro oral squamous cell carcinoma tumor tissue specimen in a fixative for fixation;

[0032] S3, and rinse with running water 0.5h before the start of the dehydration program;

[0033] S4. Fully dehydrate the washed oral squamous cell carcinoma tumor tissue specimens;

[0034] S5. Finally, the dehydrated oral squamous cell carcinoma in vitro tumor tissue specimen is placed in a solvent for transparency, and then immersed in wax for 20 minutes at a temperature of 55° C.; embedded in paraffin, sliced several times continuously, and baked to obtain the pathological sections.

[0035] In the embodiment of the present invention, the fixative in step S2 is an ethanol-formaldehyde mixed fixative, wherein the fixation time is 36 hours; the ethanol-formaldehyde mixed fixative has the functions of fixation and dehydration, which is beneficial for tumor sectioning and lymph node extraction.

[0036] In the embodiment of the present invention, the ethanol-formaldehyde mixed fixative comprises 80% ethanol and 10% formaldehyde by weight.

[0037] In the embodiment of the present invention, the solvent in step S5 is a stearic acid paraffin mixture. Typically, xylene is used as the solvent for transparency. However, xylene can easily cause tissue shrinkage, hardening, and brittleness during tissue processing. Furthermore, xylene is slightly toxic and can affect the human nervous system and mucous membranes. Therefore, a stearic acid paraffin mixture is used as the solvent. This solvent primarily contains alcohols as diluents, does not contain aromatic hydrocarbon compounds, does not produce harmful substances to the human body, does not pollute the environment, and achieves a better transparency effect.

[0038] In the embodiment of the present invention, the dehydration program in the step S4 is specific as follows: flowing water flushing 0.5h → 50% ethanol 1h → 75% ethanol 1h → 85% ethanol 2h → 95% ethanol 2h → 95% ethanol 1.5h → 100% ethanol 1.5h.At present a lot of technical personnel only pay attention to high concentration ethanol, think as long as increase the dehydration time of high concentration or directly dehydrate with 95% ethanol, just can quickly and well organize processing, yet, the result is just the opposite, and the final dehydration that handles like this is not thorough, and cell shrinkage is serious, and section quality is poor.Low ethanol penetration is strong, is easier to penetrate into tissue block inside and water is replaced out than high concentration ethanol, and contractility is little, does not cause tissue quality to change and feels that tissue is crisp when sectioning, uses 50% ethanol as the beginning of first dehydration program, though prolonged the dehydration time, achieved good effect.

[0039] In the embodiment of the present invention, during paraffin embedding in step S5, the smallest surface of the tissue in contact with the knife is placed at the bottom of the embedding cassette for longitudinal embedding, thereby facilitating subsequent sectioning operations.

[0040] In an embodiment of the present invention, in step S5, the wax block is cold-treated with an ice-water mixture before slicing. Cold-treating the wax block before slicing is the most common method, typically by placing the wax block in the freezer or directly on an ice tray. However, if fibrous or hard tissue remains in the freezer for too long, the wax block may break during rough trimming, resulting in uneven thickness and trembling slices. The uneven surface of the ice tray makes it difficult to freeze the entire wax block section uniformly, affecting the effectiveness of pretreatment. This can also cause skipping and disconnected slices during slicing. Using an ice-water mixture to pre-treat the wax block solves these problems. It maintains the appropriate hardness of the wax block and ensures uniform freezing of the entire section, saving time and effort. The resulting pathological sections are uniform, complete, free of tremor marks, wrinkles, and freshly stained. Furthermore, through better dehydration, the dehydration effect is enhanced, which in turn improves subsequent slicing results.

[0041] In an embodiment of the present invention, an ex vivo luminal squamous cell carcinoma tumor tissue specimen was immersed in an HCl solution at room temperature for 8 minutes, then immersed in trypsin-EDTA at 33°C for 15 minutes. The HCl solution concentration was 0.01N, trypsin 0.2%, and EDTA 0.008%. By treating the ex vivo luminal squamous cell carcinoma tumor tissue specimen with HCl and trypsin-EDTA, the permeability of the adipocyte membrane was increased. In this protocol, the permeability of the adipocyte membrane was enhanced through a mild chemical treatment of the tissue. However, because the lymph nodes are protected by a dense connective tissue membrane, the cell membranes of their cellular components are unaffected or minimally affected, insufficient to cause changes in cellular and tissue structure. Furthermore, the fixative used in this technical solution has the ability to emulsify and even partially dissolve fat, softening the adipose tissue surrounding and surrounding the lymph nodes, increasing the difference in hardness between the lymph nodes and the adipose tissue, making lymph node sampling easier and more convenient, and improving work efficiency and sample quality.

[0042] Example 2:

[0043] A method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma comprises the following steps:

[0044] S1. Obtain in vitro oral squamous cell carcinoma tumor tissue specimens (primary lesion specimens and lymph node specimens);

[0045] S2. Place the in vitro oral squamous cell carcinoma tumor tissue specimen in a fixative for fixation;

[0046] S3, and rinse with running water 0.75h before the start of the dehydration program;

[0047] S4. Fully dehydrate the washed oral squamous cell carcinoma tumor tissue specimens;

[0048] S5. Finally, the dehydrated oral squamous cell carcinoma in vitro tumor tissue specimen is placed in a solvent for transparency, and then immersed in wax for 22 minutes at a temperature of 62° C.; embedded in paraffin, sliced several times continuously, and baked to obtain the pathological sections.

[0049] In the embodiment of the present invention, the fixative in step S2 is an ethanol-formaldehyde mixed fixative, wherein the fixation time is 42 hours; the ethanol-formaldehyde mixed fixative has both fixation and dehydration functions, which is beneficial for tumor sectioning and lymph node extraction.

[0050] In the embodiment of the present invention, the ethanol-formaldehyde mixed fixative comprises 85% ethanol and 15% formaldehyde by weight.

[0051] In the embodiment of the present invention, the solvent in step S5 is a stearic acid paraffin mixture. Typically, xylene is used as the solvent for transparency. However, xylene can easily cause tissue shrinkage, hardening, and brittleness during tissue processing. Furthermore, xylene is slightly toxic and can affect the human nervous system and mucous membranes. Therefore, a stearic acid paraffin mixture is used as the solvent. This solvent primarily contains alcohols as diluents, does not contain aromatic hydrocarbon compounds, does not produce harmful substances to the human body, does not pollute the environment, and achieves a better transparency effect.

[0052] In the embodiment of the present invention, the dehydration program in the step S4 is specific as follows: flowing water flushing 0.5h → 50% ethanol 1h → 75% ethanol 1h → 85% ethanol 2h → 95% ethanol 2h → 95% ethanol 1.5h → 100% ethanol 1.5h.At present a lot of technical personnel only pay attention to high concentration ethanol, think as long as increase the dehydration time of high concentration or directly dehydrate with 95% ethanol, just can quickly and well organize processing, yet, the result is just the opposite, and the final dehydration that handles like this is not thorough, and cell shrinkage is serious, and section quality is poor.Low ethanol penetration is strong, is easier to penetrate into tissue block inside and water is replaced out than high concentration ethanol, and contractility is little, does not cause tissue quality to change and feels that tissue is crisp when sectioning, uses 50% ethanol as the beginning of first dehydration program, though prolonged the dehydration time, achieved good effect.

[0053] In the embodiment of the present invention, during paraffin embedding in step S5, the smallest surface of the tissue in contact with the knife is placed at the bottom of the embedding cassette for longitudinal embedding, thereby facilitating subsequent sectioning operations.

[0054] In an embodiment of the present invention, in step S5, the wax block is cold-treated with an ice-water mixture before slicing. Cold-treating the wax block before slicing is the most common method, typically by placing the wax block in the freezer or directly on an ice tray. However, if fibrous or hard tissue remains in the freezer for too long, the wax block may break during rough trimming, resulting in uneven thickness and trembling slices. The uneven surface of the ice tray makes it difficult to freeze the entire wax block section uniformly, affecting the effectiveness of pretreatment. This can also cause skipping and disjointed slices during slicing. Using an ice-water mixture to pre-treat the wax block addresses these challenges, maintaining the appropriate hardness and ensuring uniform freezing of the entire section. This saves time and effort, resulting in uniform, complete, wrinkle-free, and freshly stained pathological sections. Furthermore, through better dehydration, the dehydration effect is enhanced, simultaneously improving subsequent slicing results.

[0055] In an embodiment of the present invention, ex vivo luminal squamous cell carcinoma tumor tissue specimens were immersed in an HCl solution at room temperature for 17 minutes, then immersed in trypsin-EDTA at 35°C for 20 minutes. The HCl solution concentration was 0.03N, trypsin 1.5%, and EDTA 0.104%. Treatment of the ex vivo luminal squamous cell carcinoma tumor tissue specimens with HCl and trypsin-EDTA increased the permeability of the adipocyte membrane. In this protocol, the permeability of the adipocyte membrane was enhanced through a mild chemical treatment of the tissue. However, because the lymph nodes are protected by a dense connective tissue membrane, the cell membranes of their cellular components are unaffected or minimally affected, insufficient to cause changes in cellular and tissue structure. Furthermore, the fixative employed in this technical solution has the ability to emulsify and even partially dissolve fat, softening the adipose tissue surrounding and surrounding the lymph nodes, increasing the difference in hardness between the lymph nodes and adipose tissue, making lymph node sampling easier and more convenient, and improving both efficiency and sample quality.

[0056] Example 3:

[0057] A method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma comprises the following steps:

[0058] S1. Obtain in vitro oral squamous cell carcinoma tumor tissue specimens (primary lesion specimens and lymph node specimens);

[0059] S2. Place the in vitro oral squamous cell carcinoma tumor tissue specimen in a fixative for fixation;

[0060] S3, and rinse with running water 1 hour before the start of the dehydration program;

[0061] S4. Fully dehydrate the washed oral squamous cell carcinoma tumor tissue specimens;

[0062] S5. Finally, the dehydrated oral squamous cell carcinoma in vitro tumor tissue specimen is placed in a solvent for transparency, and then immersed in wax for 25 minutes at a temperature of 70° C.; embedded in paraffin, sliced several times continuously, and baked to obtain the pathological sections.

[0063] In the embodiment of the present invention, the fixative in step S2 is an ethanol-formaldehyde mixed fixative, wherein the fixation time is 348 hours; the ethanol-formaldehyde mixed fixative has the functions of fixation and dehydration, which is beneficial for tumor sectioning and lymph node extraction.

[0064] In the embodiment of the present invention, the ethanol-formaldehyde mixed fixative comprises 90% ethanol and 20% formaldehyde by weight.

[0065] In the embodiment of the present invention, the solvent in step S5 is a stearic acid paraffin mixture. Typically, xylene is used as the solvent for transparency. However, xylene can easily cause tissue shrinkage, hardening, and brittleness during tissue processing. Furthermore, xylene is slightly toxic and can affect the human nervous system and mucous membranes. Therefore, a stearic acid paraffin mixture is used as the solvent. This solvent primarily contains alcohols as diluents, does not contain aromatic hydrocarbon compounds, does not produce harmful substances to the human body, does not pollute the environment, and achieves a better transparency effect.

[0066] In the embodiment of the present invention, the dehydration program in the step S4 is specific as follows: flowing water flushing 0.5h → 50% ethanol 1h → 75% ethanol 1h → 85% ethanol 2h → 95% ethanol 2h → 95% ethanol 1.5h → 100% ethanol 1.5h.At present a lot of technical personnel only pay attention to high concentration ethanol, think as long as increase the dehydration time of high concentration or directly dehydrate with 95% ethanol, just can quickly and well organize processing, yet, the result is just the opposite, and the final dehydration that handles like this is not thorough, and cell shrinkage is serious, and section quality is poor.Low ethanol penetration is strong, is easier to penetrate into tissue block inside and water is replaced out than high concentration ethanol, and contractility is little, does not cause tissue quality to change and feels that tissue is crisp when sectioning, uses 50% ethanol as the beginning of first dehydration program, though prolonged the dehydration time, achieved good effect.

[0067] In the embodiment of the present invention, during paraffin embedding in step S5, the smallest surface of the tissue in contact with the knife is placed at the bottom of the embedding cassette for longitudinal embedding, thereby facilitating subsequent sectioning operations.

[0068] In an embodiment of the present invention, in step S5, the wax block is cold-treated with an ice-water mixture before slicing. Cold-treating the wax block before slicing is the most common method, typically by placing the wax block in the freezer or directly on an ice tray. However, if fibrous or hard tissue remains in the freezer for too long, the wax block may break during rough trimming, resulting in uneven thickness and trembling slices. The uneven surface of the ice tray makes it difficult to freeze the entire wax block section uniformly, affecting the effectiveness of pretreatment. This can also cause skipping and disjointed slices during slicing. Using an ice-water mixture to pre-treat the wax block addresses these challenges, maintaining the appropriate hardness and ensuring uniform freezing of the entire section. This saves time and effort, resulting in uniform, complete, wrinkle-free, and freshly stained pathological sections. Furthermore, through better dehydration, the dehydration effect is enhanced, simultaneously improving subsequent slicing results.

[0069] In an embodiment of the present invention, an ex vivo luminal squamous cell carcinoma tumor tissue specimen was immersed in an HCl solution at room temperature for 25 minutes, then immersed in trypsin-EDTA at 38°C for 25 minutes. The HCl solution concentration was 0.05N, trypsin 2.8%, and EDTA 0.2%. By treating the ex vivo luminal squamous cell carcinoma tumor tissue specimen with HCl and trypsin-EDTA, the permeability of the adipocyte membrane was increased. In this protocol, the permeability of the adipocyte membrane was enhanced through a mild chemical treatment of the tissue. However, because the lymph nodes are protected by a dense connective tissue membrane, the cell membranes of their cellular components are unaffected or only slightly affected, insufficient to cause changes in cellular and tissue structure. Furthermore, the fixative used in this technical solution has the effect of emulsifying and even partially dissolving fat, softening the adipose tissue surrounding and surrounding the lymph nodes, increasing the difference in hardness between the lymph nodes and the adipose tissue, making lymph node sampling easier and more convenient, and improving work efficiency and sample quality.

[0070] Meanwhile, the contents not described in detail in this specification belong to the prior art known to those skilled in the art.

[0071] It should be noted that, in this document, relational terms such as first and second, etc., are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus.

[0072] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma, characterized in that: The specific steps include: S1. Obtain in vitro tumor tissue specimens of oral squamous cell carcinoma; S2. Place the in vitro oral squamous cell carcinoma tumor tissue specimen in a fixative for fixation; S3, and rinse with running water 0.5 to 1 hour before the start of the dehydration program; S4. Fully dehydrate the washed oral squamous cell carcinoma tumor tissue specimens; S5. Finally, the dehydrated oral squamous cell carcinoma in vitro tumor tissue specimen is placed in a solvent for transparency, and then immersed in wax, embedded in paraffin, sliced several times continuously, and baked to obtain the pathological sections.

2. The method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma according to claim 1, characterized in that: The oral squamous cell carcinoma in vitro tumor tissue specimens in step S1 include primary lesion specimens and lymph node specimens.

3. The method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma according to claim 1, characterized in that: The fixative in step S2 is an ethanol-formaldehyde mixed fixative, wherein the fixative time is 36 to 48 hours.

4. The method for processing an in vitro oral squamous cell carcinoma tumor tissue specimen according to claim 3, characterized in that: According to the mass ratio, the ethanol-formaldehyde mixed fixative contains 80-90% ethanol and 10-20% formaldehyde.

5. The method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma according to claim 1, characterized in that: In step S5, the wax immersion time is 20 to 25 minutes, the wax immersion temperature is 55 to 70° C., and the solvent is a stearic acid paraffin mixture.

6. The method for processing an in vitro oral squamous cell carcinoma tumor tissue specimen according to claim 1, characterized in that: The dehydration procedure in step S4 is as follows: running water washing for 0.5 h → 50% ethanol for 1 h → 75% ethanol for 1 h → 85% ethanol for 2 h → 95% ethanol for 2 h → 95% ethanol for 1.5 h → 100% ethanol for 1.5 h.

7. The method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma according to claim 1, characterized in that: In step S5, when embedding in paraffin, the smallest surface of the tissue in contact with the knife is placed at the bottom of the embedding box for longitudinal embedding.

8. The method for processing an in vitro tumor tissue specimen of oral squamous cell carcinoma according to claim 1, characterized in that: In step S5, before sectioning begins, the wax block is cooled using an ice-water mixture.

9. The method for processing an in vitro oral squamous cell carcinoma tumor tissue specimen according to claim 1, characterized in that: The method for processing an oral squamous cell carcinoma in vitro tumor tissue specimen also includes pre-processing the tissue specimen: immersing the obtained oral squamous cell carcinoma in vitro tumor tissue specimen in HCl solution at room temperature for 8 to 25 minutes, and then immersing it in trypsin-EDTA at a constant temperature of 33 to 38°C for 15 to 25 minutes.

10. Use of an in vitro oral squamous cell carcinoma tumor tissue specimen treated according to any one of claims 1 to 9 in a routine pathological section examination.

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