A degreasing liquid for naked eye visualization of surgical pathological specimens and a preparation method thereof

Abstract

The present invention provides a kind of degreasing liquid and preparation method thereof for naked eye visualization of surgical pathology specimens, including 2-20 weight parts of permeabilizing agent, 30-60 weight parts of penetration enhancer, 40-70 weight parts of degreasing agent and 0.1-2 weight parts of solubilizing agent. Wherein, permeabilizing agent is methyl, ethyl substituted naphthalene compound; penetration enhancer is C1-C3 lower alcohol; degreasing agent is C6-C8 cycloalkane; solubilizing agent is polyethylene glycol type nonionic surfactant. The present invention solves the problem of accurate acquisition of parenchyma tissue such as nerve, blood vessel, lymph node, cancerous nodule, organ stump, etc. in adipose tissue (including but not limited to omentum, stomach periphery, intestinal periphery, pancreatic periphery, axilla, liver hilum, renal hilum adipose tissue, etc.) in existing pathology specimens, in subsequent pathology film making, all parenchyma tissues obtained retain their original tissue structure, without affecting morphological observation, immunohistochemistry and in situ hybridization effect.

Description

Technical Field

[0001] The invention belongs to the technical field of pathological specimen processing, and relates to a pathological specimen processing liquid, and in particular to a defatting liquid for naked eye visualization processing of surgical pathological specimens, and a preparation method and application thereof. Background Art

[0002] Distant metastasis is one of the important biological characteristics of malignant tumors and an important basis for clinical TNM staging. There are four ways of metastasis: ① Direct spread to adjacent sites, the most common of which is invasion of nerve tissue around visceral organs, such as invasion of brachial plexus nerves by apical lung tumors, resulting in shoulder and arm pain; ② Hematogenous metastasis: cancer cells invade blood vessels and metastasize to distant sites such as lungs, liver, bones, brain, etc. with blood flow, forming secondary tumors; ③ Lymphatic metastasis: cancer cells in the primary site are transferred to lymph nodes at all levels from near to far with lymphatic drainage, and may also undergo skipping metastasis; ④ Implantation: cancer cells are implanted in pleural or peritoneal fat tissue after detachment. Radical surgery of R0 complete resection of the primary lesion and normal tissue within a certain range around it and clearing of drainage regional lymph nodes is an important surgical method in clinical practice, and the standardized specimens of surgical resection specimens sent to the pathology department after surgery (especially the judgment of whether there is distant metastasis) are one of the important steps to ensure the quality of pathology reports, infer patient prognosis, and guide clinical follow-up treatment.

[0003] Adequate pathological specimens can maximize the detection rate of risk factors that lead to cancer-related adverse events (such as nerve invasion, intravascular cancer thrombus, lymph node metastasis, and peritoneal metastasis), and also have an important impact on the prediction of patients' long-term survival rate. Studies have confirmed that more than half of pancreatic ductal adenocarcinomas have lymph node metastasis and nerve invasion. Clinically, extensive retroperitoneal soft tissue resection and expanded lymph node dissection are advocated to achieve a better prognosis. Early microvascular invasion and the formation of portal vein cancer thrombus in the middle and late stages are the most important factors affecting the poor prognosis of hepatocellular carcinoma after surgery. Lymph node metastasis staging (N staging) based on the number and anatomical location of lymph node metastasis is the most common prognostic evaluation and diagnostic staging method for gastrointestinal and breast tumors. Many special types of lymph node metastasis can also be confirmed through sufficient specimens, such as cancer nodules, lymph node micrometastasis, isolated tumor cells and skip metastasis, thereby reducing the incidence of false negative lymph nodes. Patients with colorectal cancer and peritoneal metastasis who undergo radical resection have a better prognosis than those with synchronous liver metastasis. Therefore, localized colorectal cancer with peritoneal metastasis may be cured by surgery. In summary, sufficient pathological specimens are of great significance for determining the stage of malignant tumors, evaluating prognosis, and formulating individualized treatment plans.

[0004] The sampling of clinical malignant tumor radical specimens is the focus and difficulty of pathological work. The objects mentioned above that need to be fully sampled and carefully observed under a microscope (nerves, blood vessels, lymph nodes, cancer nodules, etc.) are all buried in adipose tissue. Pathologists often use a method of cutting with a knife at a certain distance and touching it by hand. According to the experience of hand feeling, they judge the parenchymal tissue wrapped in adipose tissue and sample it. There are great randomness and human operation factors, especially when the adipose tissue is too abundant and the volume of the parenchymal tissue is less than 0.5cm in diameter. In this case, the possibility of micro-metastatic cancer lesions is often missed. The current solution can only be found patiently. It may take several hours to fully sample the key observation object for a radical specimen, which suddenly increases time and labor costs. Therefore, in the actual work of the pathology department, there is an urgent need for a method to efficiently and quickly visualize adipose tissue of various sizes with the naked eye, reveal the parenchymal tissues such as nerves, blood vessels, lymph nodes, etc. contained therein, and fully fix them.

[0005] Literature search revealed that in recent years, many studies have focused on the use of fat-dissolving solutions to treat breast cancer radical resection specimens to improve the detection rate of axillary lymph nodes, but have not mentioned the effects on other important parenchymal tissues (nerves, blood vessels, lymph nodes, cancer nodules, etc.), and there have been no reports on the use of naked eye visualization processing methods for large specimens and small biopsy specimens containing adipose tissue after radical resection of common clinical malignant tumors including the gastrointestinal tract, pancreas, liver, kidneys and other organs. Summary of the invention

[0006] The present invention is based on the above research. In view of the difficulties existing in the sample of tiny parenchymal tissue in the existing adipose tissue specimens and the processing defects of the current fat dissolving solution, a new degreasing solution for naked eye visualization is provided. The degreasing solution has the multiple functions of transparency, fat removal, and fixation of various parenchymal tissues (nerves, blood vessels, lymph nodes, cancer nodules, organ stumps, etc.) in adipose tissue. While transparently removing adipose tissue, it can effectively retain the morphological structure, immunohistochemical antigen epitopes and in situ hybridization nucleic acid sequences of various parenchymal tissues.

[0007] In order to achieve this purpose, the technical solution adopted by the present invention is as follows:

[0008] The first aspect of the present invention provides a degreasing solution for naked eye visualization of surgical pathological specimens, which is composed of four components: a permeabilizing agent, a penetration enhancer, a degreasing agent, and a solubilizing agent. The weight parts of the four components are as follows: the permeabilizing agent is a methyl or ethyl substituted naphthalene compound, 2-20 parts by weight; the penetration enhancer is a C1-C3 lower alcohol, 30-60 parts by weight; the degreasing agent is a C6-C8 cycloalkane, 40-70 parts by weight; and the solubilizing agent is a polyethylene glycol-type nonionic surfactant, 0.1-2 parts by weight.

[0009] Preferably, the permeabilizing agent is 5-15 parts by weight, the penetration enhancer is 40-55 parts by weight, the degreasing agent is 45-65 parts by weight and the solubilizing agent is 0.2-1 parts by weight.

[0010] Further preferably, the specific reagents of the four components are selected as follows:

[0011] The permeabilizing agent is one or a mixture of methylnaphthalene, ethylnaphthalene, dimethylnaphthalene, diethylnaphthalene and trimethylnaphthalene;

[0012] The penetration enhancer is one or a mixture of methanol, ethanol, propanol, and isopropanol;

[0013] The degreasing agent is one or a mixture of cyclohexane, methylcyclohexane, and ethylcyclohexane;

[0014] The solubilizer is one or a mixture of polyethylene glycol monooctylphenyl ether, polyethylene glycol monononylphenyl ether, fatty alcohol polyoxyethylene ether, and polyoxyethylene castor oil.

[0015] More preferably, the specific composition of the degreasing liquid is as described in any one of the following (1) to (5):

[0016] (1) the degreasing liquid is composed of 9.5 parts by weight of 1-methylnaphthalene or 2-methylnaphthalene, 45 parts by weight of ethanol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of polyethylene glycol monooctylphenyl ether;

[0017] (2) the degreasing liquid is composed of 12.5 parts by weight of 1-ethylnaphthalene or 2-ethylnaphthalene, 40 parts by weight of methanol, 47 parts by weight of methylcyclohexane, and 0.5 parts by weight of polyethylene glycol monononylphenyl ether;

[0018] (3) the degreasing liquid is composed of 10 parts by weight of 1,2-dimethylnaphthalene, 1,4-dimethylnaphthalene, 1,5-dimethylnaphthalene or 2,7-dimethylnaphthalene, 42 parts by weight of propanol, 47 parts by weight of ethylcyclohexane and 1 part by weight of fatty alcohol polyoxyethylene ether;

[0019] (4) the degreasing liquid is composed of 8.5 parts by weight of 2,6-diethylnaphthalene, 45 parts by weight of isopropanol, 46 parts by weight of cyclohexane, and 0.5 parts by weight of polyoxyethylene castor oil;

[0020] (5) The degreasing liquid is composed of 9.5 parts by weight of 1,2,4-trimethylnaphthalene, 1,4,6-trimethylnaphthalene, 2,3,5-trimethylnaphthalene or 2,3,6-trimethylnaphthalene, 45 parts by weight of ethanol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of polyethylene glycol monooctylphenyl ether.

[0021] The second aspect of the present invention provides a method for preparing the above-mentioned degreasing liquid, comprising the following steps: weighing a permeabilizing agent, a penetration enhancer, a degreasing agent and a solubilizing agent according to selected weight portions, and blending and mixing them at room temperature to 45°C to obtain the degreasing liquid.

[0022] The third aspect of the present invention provides the use of the above-mentioned degreasing liquid in the degreasing of surgical pathological specimens.

[0023] Preferably, the surgical pathological specimens include surgical gross specimens or biopsy tissues. The purpose is to defat the surgical gross specimens or biopsy tissues and visualize the non-fat tissue inside them with the naked eye, and the non-fat tissue includes parenchymal tissues such as nerves, blood vessels, lymph nodes, cancerous nodules, and organ stumps.

[0024] In a fourth aspect of the present invention, a method for visualizing surgical pathological specimens using the above-mentioned defatting liquid is provided: a surgical gross specimen or a biopsy small tissue is placed in a defatting liquid at a weight-to-volume ratio of 1:3-1:100 at room temperature, and the specimen is immersed in the defatting liquid for more than 2 hours, during which the liquid can be changed and the immersion can be repeated. Subsequently, specific parts of the pathological specimen can be sampled and obtained under the naked eye for pathological slide preparation.

[0025] Through experiments, it is verified that the degreasing liquid of the present invention can be used to process gross surgical specimens or small biopsy tissues at the same time, and has the functions of transparentizing, degreasing, and fixing specimens. Compared with the currently commonly used formaldehyde treatment liquid, it can significantly improve the detection rate of non-fat tissue in samples without affecting the subsequent morphological observation, immunohistochemistry and in situ hybridization effects. It is an ideal formula reagent that can be used for pathological specimens of adipose tissue of different sizes.

[0026] Beneficial protection and effects of the present invention:

[0027] When the degreasing liquid of the present invention is applied to gross specimens or small biopsy tissues of conventional radical surgery for malignant tumors, it can effectively make fat tissues of different sizes transparent, and also has a certain fat removal function, and ensures sufficient fixation of the inner substantial tissues.

[0028] Because this defatting liquid has the function of transparent fat tissue and makes it visible to the naked eye, it improves the detection rate of hidden solid tissue (nerves, blood vessels, lymph nodes, cancer nodules, organ stumps, etc.), and thus accurately provides evidence of distant metastasis of malignant tumors (nerve invasion, intravascular cancer thrombus, lymph node metastasis, cancer nodules, etc.), saving the time spent by pathologists on sample inspection, greatly improving labor efficiency, and saving labor costs, which is of great practical significance, especially in the current situation of an extreme shortage of pathologists.

[0029] Experiments have confirmed that in the subsequent pathological preparation of surgical gross specimens or biopsy small tissues treated with this defatting solution, all the solid tissues obtained retain their original tissue structure, and do not affect the morphological observation, immunohistochemistry and in situ hybridization effects.

[0030] Therefore, the present invention solves the problem of accurate acquisition of parenchymal tissues such as nerves, blood vessels, lymph nodes, cancerous nodules, organ stumps, etc. in adipose tissue (including but not limited to omentum, perigastric, periintestinal, peripancreatic, axillary, hepatic hilar, renal hilar adipose tissue, etc.) in existing pathological specimens. The degreasing liquid of the present invention is promoted and used in pathological specimens after commercialization, which can generate certain economic value, and its material collection and preparation process are simple, which can be configured by the laboratory itself and is conducive to industrial scale expansion of production. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 To observe the transparency of fat tissue in the control group and the test group (Examples 1, 3, 5, 9, 10) by naked eye comparison;

[0032] Figure 2 The light transmittance of adipose tissue treated with different defatting solutions in the control group and the test group (Examples 1 to 13) was measured using a LS116 light transmittance meter;

[0033] Figure 3 Comparison of paraffin sections and immunohistochemical microscopic pathological images of nerve tissue in the adipose tissue of the liver hilum in the control group and the experimental group (Examples 1, 3, 5, 9, 10);

[0034] Figure 4 Comparison of paraffin sections and immunohistochemical microscopic pathological images of vascular tissue in the hepatic hilar cleaned adipose tissue of the control group and the experimental group (Examples 1, 3, 5, 9, 10);

[0035] Figure 5 Comparison of paraffin sections, immunohistochemistry and in situ hybridization sections of lymph nodes in the adipose tissue of the liver hilum in the control group and the experimental group (Example 1);

[0036] Figure 6 Comparison of paraffin sections and immunohistochemical sections of cancer nodules in adipose tissue of the liver hilum in the control group and the experimental group (Examples 1, 3, 5, 9, 10) under microscopic pathology;

[0037] Figure 7 Comparison of paraffin sections and immunohistochemical sections of organ stumps (part of colon or pancreatic tissue) in adipose tissue of the control group and the experimental group (Examples 1, 3, 5, 9, 10) after hepatic hilar cleanup and pathological images of immunohistochemical sections;

[0038] Figure 8 Comparison of paraffin sections and immunohistochemical sections of abdominal nodules in the control group and the experimental group (Examples 1, 3, 5, 9, 10) under microscopic pathology. DETAILED DESCRIPTION

[0039] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0040] The experimental methods in the following examples without specifying specific conditions are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are calculated by weight. Unless otherwise defined, all professional and scientific terms used in the text have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described can be applied to the present invention. The preferred implementation methods and materials described in the text are for demonstration purposes only.

[0041] 1. The formula of the degreasing solution for naked eye visualization of surgical pathological specimens is shown in Examples 1 to 13:

[0042] Example 1

[0043] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on 100% total weight, 9.5% 1-methylnaphthalene, 45% ethanol, 45% cyclohexane, and 0.5% polyethylene glycol monooctylphenyl ether.

[0044] Example 2

[0045] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on a total weight of 100%, 9.5% of 2-methylnaphthalene, 45% of ethanol, 45% of cyclohexane, and 0.5% of polyethylene glycol monooctylphenyl ether.

[0046] Example 3

[0047] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on 100% total weight, 12.5% ​​1-ethylnaphthalene, 40% methanol, 47% methylcyclohexane, and 0.5% polyethylene glycol monononylphenyl ether.

[0048] Example 4

[0049] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on 100% total weight, 12.5% ​​2-ethylnaphthalene, 40% methanol, 47% methylcyclohexane, and 0.5% polyethylene glycol monononylphenyl ether.

[0050] Example 5

[0051] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on a total weight of 100%, 1,2-dimethylnaphthalene 10%, propanol 42%, ethylcyclohexane 47%, and fatty alcohol polyoxyethylene ether 1%.

[0052] Example 6

[0053] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on a total weight of 100%, 1,4-dimethylnaphthalene 10%, propanol 42%, ethylcyclohexane 47%, and fatty alcohol polyoxyethylene ether 1%.

[0054] Example 7

[0055] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on 100% total weight, 10% 1,5-dimethylnaphthalene, 42% propanol, 47% ethylcyclohexane, and 1% fatty alcohol polyoxyethylene ether.

[0056] Example 8

[0057] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on 100% total weight, 10% 2,7-dimethylnaphthalene, 42% propanol, 47% ethylcyclohexane, and 1% fatty alcohol polyoxyethylene ether.

[0058] Example 9

[0059] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on 100% total weight, 8.5% 2,6-diethylnaphthalene, 45% isopropanol, 46% cyclohexane, and 0.5% polyoxyethylene castor oil.

[0060] Example 10

[0061] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on a total weight of 100%, 9.5% of 1,2,4-trimethylnaphthalene, 45% of ethanol, 45% of cyclohexane, and 0.5% of polyethylene glycol monooctylphenyl ether.

[0062] Embodiment 11

[0063] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on a total weight of 100%, 9.5% of 1,4,6-trimethylnaphthalene, 45% of ethanol, 45% of cyclohexane, and 0.5% of polyethylene glycol monooctylphenyl ether.

[0064] Example 12

[0065] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on a total weight of 100%, 9.5% of 2,3,5-trimethylnaphthalene, 45% of ethanol, 45% of cyclohexane, and 0.5% of polyethylene glycol monooctylphenyl ether.

[0066] Example 13

[0067] The degreasing liquid used for naked eye visualization of surgical pathological specimens in this embodiment includes, based on a total weight of 100%, 9.5% of 2,3,6-trimethylnaphthalene, 45% of ethanol, 45% of cyclohexane, and 0.5% of polyethylene glycol monooctylphenyl ether.

[0068] The degreasing solution preparation methods in the above embodiments are the same, and all adopt the following preparation method: the above permeabilizing agent, penetration enhancer, degreasing agent, and solubilizing agent are blended at room temperature to 45° C. to form the degreasing solution for naked eye visualization of surgical pathological specimens.

[0069] The above formulas are analyzed and classified according to the permeabilizing agent: Examples 1 and 2 both belong to the monosubstituted methylnaphthalene system; Examples 3 and 4 both belong to the monosubstituted ethylnaphthalene system; Examples 5 to 8 all belong to the dimethylnaphthalene system; Example 9 belongs to the diethylnaphthalene system; Examples 10 to 13 all belong to the trimethylnaphthalene system.

[0070] The performance of the permeabilizing agents in the same system is similar. Hereinafter, one degreasing liquid is selected from each of the above five systems for experimental verification, and the degreasing liquids shown in Example 1, Example 3, Example 5, Example 9 and Example 10 are selected as representatives for subsequent experiments.

[0071] 2. Effect Verification

[0072] The following experimental examples are used to better illustrate the transparent, fat-removing and fixation effects of the degreasing solution of the present invention on adipose tissue. Compared with adipose tissue not treated with the degreasing solution, the fixation and preservation effects are the same, but the light transmittance and transparency of the adipose tissue are greatly enhanced, which is more conducive to the exposure of the parenchymal tissue inside and the acquisition of the sample, and can achieve the significant technical effect of effectively retaining the morphological structure, immunohistochemical antigen epitopes and in situ hybridization nucleic acid sequences of the tissue to be tested.

[0073] 1. Experimental materials and reagents

[0074] Experimental materials: Hepatic hilar adipose tissue (gross surgical specimens) or abdominal nodules (small biopsy tissues).

[0075] Experimental reagents: The degreasing solution of the test group was the degreasing solution prepared according to the above example, and the control group was the traditional 4% neutral formaldehyde fixative.

[0076] 2. Experimental methods and results

[0077] 2.1 Experimental methods

[0078] The hepatic hilar cleaned adipose tissue or abdominal fat was immersed in the solution of the experimental group and the control group at a weight-to-volume ratio of 1:3-1:100 at room temperature for 6 hours. If necessary, the solution was changed and the immersion was repeated. Subsequently, the specific parts of the pathological specimens were sampled and obtained under the naked eye for routine pathological preparation, immunohistochemical staining and in situ hybridization.

[0079] 2.2 Transparency and transmittance comparison

[0080] The transparency of the fat tissue in the control group and the experimental group was observed by naked eyes. The transmittance of the two groups was measured using a LS116 transmittance meter. The transmittance measurement method is as follows:

[0081] Transparency measurement of control group:

[0082] (1) Take the fixed tissue, flatten it and place it in a 4°C refrigerator for 12 hours. After the fat tissue hardens, cut it into 4 cm × 4 cm tissue blocks with a thickness of about 5 mm;

[0083] (2) The tissue block was spread in a large quartz transparent cuvette (5 cm × 5 cm × 1 cm) and a 50% ethanol aqueous solution was added;

[0084] (3) The transmittance (%) was measured using a LS116 transmittance meter. This is the transmittance (%) of the adipose tissue in the control group.

[0085] Determination of transparency of processed tissue:

[0086] (1) Take the fixed tissue, flatten it and place it in a 4°C refrigerator for 12 hours. After the fat tissue hardens, cut it into 4 cm × 4 cm tissue blocks with a thickness of about 5 mm;

[0087] (2) The tissue block was immersed in the degreasing solution prepared in Example 1-13 at room temperature for 6 hours;

[0088] (3) Take out the tissue block, spread it in a large quartz transparent cuvette (5 cm × 5 cm × 1 cm), and add 50% ethanol aqueous solution;

[0089] (4) The transmittance (%) was measured using a LS116 transmittance meter. This is the transmittance (%) of the adipose tissue in the experimental group.

[0090] The results of visually comparing the transparency of the fat tissue in the control group and the test group (Examples 1, 3, 5, 9, 10) are shown in Figure 1 ; The light transmittance test comparison results are shown in Table 1 and Figure 2 The results show that under the same photographing conditions, the transparency of the samples treated with the degreasing solution of Examples 1, 3, 5, 9, and 10 of the present invention is significantly better than that of the control group ( Figure 1 ); The transmittance test results show that the transparency of the sample treated with the degreasing solution of the present invention is 8 to 10 times that of the control group sample test data, and the difference is significant (Table 1 and Figure 2 ).

[0091] Table 1 Comparison of light transmittance between the experimental group and the control group

[0092] Group Light transmittance (%) OP_SD Control group 3.2 0.45 Example 1 28.8 0.50 Example 2 27.1 2.00 Example 3 32.5 1.83 Example 4 23.6 3.68 Example 5 24.4 3.91 Example 6 26.2 1.96 Example 7 22.7 3.43 Example 8 24.9 2.56 Example 9 23.6 3.61 Example 10 25.3 3.20 Embodiment 11 26.4 4.82 Example 12 22.5 3.67 Example 13 23.8 2.32

[0093] 2.3 Comparison of immunohistochemistry and in situ hybridization

[0094] The adipose tissue of the experimental group and the control group was taken out from the defatting solution or fixative solution, and the solid tissues in it were sampled one by one and placed in the embedding box. The subsequent dehydration, transparency, wax immersion, embedding, sectioning, immunohistochemistry and in situ hybridization of the above solid tissues were carried out according to the standardized process. The results are as follows:

[0095] 2.3.1 Morphological observation and immunohistochemical effect of nerve tissue in adipose tissue removed from the hepatic hilum

[0096] The paraffin sections / immunohistochemical sections of the nerve tissue in the hepatic hilar adipose tissue prepared in the test groups (Examples 1, 3, 5, 9, 10) were compared with the paraffin sections / immunohistochemical sections of the nerve tissue in the hepatic hilar adipose tissue prepared in the control group. Figure 3 ,Comparison found that the morphological observation and immunohistochemical effects of the experimental group and the control group were basically the same.

[0097] 2.3.2 Morphological observation and immunohistochemical effect of vascular tissue in adipose tissue removed from the hepatic hilum

[0098] The paraffin sections / immunohistochemical sections of vascular tissue in the hepatic hilar adipose tissue cleared from the experimental groups (Examples 1, 3, 5, 9, 10) were compared with the paraffin sections / immunohistochemical sections of vascular tissue in the hepatic hilar adipose tissue cleared from the control group. Figure 4 ,Comparison found that the morphological observation and immunohistochemical effects of the experimental group and the control group were basically the same.

[0099] 2.3.3 Morphological observation and immunohistochemical effect of lymph nodes in adipose tissue removed from the hepatic hilum

[0100] The paraffin sections / immunohistochemical sections / in situ hybridization of lymph nodes in the dissected adipose tissue of the hepatic hilar region prepared in the test group (Example 1) were compared with the paraffin sections / immunohistochemical sections / in situ hybridization sections of lymph nodes in the dissected adipose tissue of the hepatic hilar region prepared in the control group. The results are shown in Figure 5 ,Comparison showed that the morphological observation, immunohistochemistry and in situ hybridization effects of the experimental group and the control group were basically the same.

[0101] 2.3.4 Morphological observation and immunohistochemical effect of clearing cancer nodules in adipose tissue at the hepatic hilum

[0102] The paraffin sections / immunohistochemical sections of the cancer nodules in the adipose tissue of the hepatic hilar region prepared in the test groups (Examples 1, 3, 5, 9, 10) were compared with the paraffin sections / immunohistochemical sections of the cancer nodules in the adipose tissue of the hepatic hilar region prepared in the control group. Figure 6,Comparison found that the morphological observation and immunohistochemical effects of the experimental group and the control group were basically the same.

[0103] 2.3.5 Morphological observation and immunohistochemical results of organ stumps in adipose tissue cleared from the hepatic portal (partial intestinal wall tissue or pancreatic tissue)

[0104] The paraffin sections / immunohistochemical sections of the organ stumps in the adipose tissue of the hepatic hilar region prepared in the test groups (Examples 1, 3, 5, 9, 10) were compared with the paraffin sections / immunohistochemical sections of the organ stumps in the adipose tissue of the hepatic hilar region prepared in the control group. Figure 7 ,Comparison found that the morphological observation and immunohistochemical effects of the experimental group and the control group were basically the same.

[0105] 2.3.6 Morphological observation and immunohistochemical effect on abdominal nodules

[0106] The paraffin sections / immunohistochemical sections of the abdominal nodules prepared in the test groups (Examples 1, 3, 5, 9, 10) were compared with the paraffin sections / immunohistochemical sections of the abdominal nodules prepared in the control group. Figure 8 ,Comparison found that the morphological observation and immunohistochemical effects of the experimental group and the control group were basically the same.

[0107] 2.3.7 Conclusion

[0108] The degreasing liquid for naked eye visualization of surgical pathological specimens of the present invention can be used to process gross surgical specimens or small biopsy tissues, and has the functions of transparency, fat removal, and specimen fixation, and does not affect subsequent morphological observation, immunohistochemistry, and in situ hybridization effects. It is an ideal formula reagent that can be used for pathological specimens of adipose tissue of different sizes.

[0109] The examples of the present invention have been specifically described above, but the present invention is not limited to the embodiments. Those skilled in the art may make various equivalent modifications or substitutions without violating the creative spirit of the present invention. These equivalent modifications or substitutions are all included in the scope defined by the claims of this application.

Claims

1. A degreasing solution for naked eye visualization of surgical pathological specimens, characterized in that: Based on the total weight of 100%, it is composed of four components: a permeation agent, a penetration enhancer, a degreasing agent, and a solubilizing agent. The weight proportions of the four components are as follows: Wherein, the permeabilizing agent is one of methylnaphthalene, ethylnaphthalene, dimethylnaphthalene, diethylnaphthalene and trimethylnaphthalene, 8-12.5 parts by weight; The penetration enhancer is one of methanol, ethanol, propanol and isopropanol, 40-45 parts by weight; The degreasing agent is one of cyclohexane, methylcyclohexane and ethylcyclohexane, 45-47 parts by weight; The solubilizing agent is one of polyethylene glycol monooctylphenyl ether, polyethylene glycol monononylphenyl ether, fatty alcohol polyoxyethylene ether, and polyoxyethylene castor oil, with a weight portion of 0.5-1.

2. The degreasing solution for naked eye visualization of surgical pathological specimens according to claim 1, characterized in that: in, The composition of the degreasing liquid is as described in any one of the following (1) to (5): (1) The degreasing liquid is composed of 9.5 parts by weight of 1-methylnaphthalene or 2-methylnaphthalene, 45 parts by weight of ethanol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of polyethylene glycol monooctylphenyl ether; (2) the degreasing liquid is composed of 12.5 parts by weight of 1-ethylnaphthalene or 2-ethylnaphthalene, 40 parts by weight of methanol, 47 parts by weight of methylcyclohexane, and 0.5 parts by weight of polyethylene glycol monononylphenyl ether; (3) The degreasing liquid is composed of 10 parts by weight of 1,2-dimethylnaphthalene, 1,4-dimethylnaphthalene, 1,5-dimethylnaphthalene or 2,7-dimethylnaphthalene, 42 parts by weight of propanol, 47 parts by weight of ethylcyclohexane and 1 part by weight of fatty alcohol polyoxyethylene ether; (4) the degreasing liquid is composed of 8.5 parts by weight of 2,6-diethylnaphthalene, 45 parts by weight of isopropanol, 46 parts by weight of cyclohexane, and 0.5 parts by weight of polyoxyethylene castor oil; (5) The degreasing liquid is composed of 9.5 parts by weight of 1,2,4-trimethylnaphthalene, 1,4,6-trimethylnaphthalene, 2,3,5-trimethylnaphthalene or 2,3,6-trimethylnaphthalene, 45 parts by weight of ethanol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of polyethylene glycol monooctylphenyl ether.

3. The degreasing solution for naked eye visualization of surgical pathological specimens according to claim 1, characterized in that: in, The surgical pathological specimens include gross surgical specimens or small biopsy tissues.

4. The method for preparing the degreasing solution for naked eye visualization of surgical pathological specimens according to claim 1, characterized in that: The steps include: The permeabilizing agent, penetration enhancer, degreasing agent and solubilizing agent are weighed according to the selected weight portions, and blended and mixed at room temperature to 45°C.

5. Use of the degreasing solution according to any one of claims 1 to 2 in transparentizing adipose tissue of surgical pathological specimens.

6. The use according to claim 5, characterized in that: The purpose is to defat surgical gross specimens or biopsy tissues and visualize the non-fat tissue inside them with the naked eye, wherein the non-fat tissue includes nerves, blood vessels, lymph nodes, cancerous nodules, and organ stumps.

7. A method for visualizing surgical pathology specimens using the degreasing solution according to any one of claims 1 to 2, characterized in that: Place the surgical gross specimen or small biopsy tissue in the degreasing solution at a weight-to-volume ratio of 1:3-1:100 at room temperature, and soak and defat for more than 2 hours. During this period, the solution can be changed and the soaking can be repeated.

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