A degreasing solution for permeabilizing surgical specimens and a preparation method thereof

Abstract

The present invention provides a degreasing solution for permeabilization of surgical gross specimens and a preparation method thereof, which is composed of four components: a permeabilizer, a penetration enhancer, a degreasing agent, and a solubilizer, and the weight proportions of the four components are as follows: the permeabilizer is a five-membered or six-membered azacyclic ketone compound, and its N-position substituent is a methyl, ethyl or butyl group, with a weight proportion of 2-20%; the penetration enhancer is a C1-C3 lower alcohol, with a weight proportion of 30%-55%; the degreasing agent is a C6-C8 cycloalkane, with a weight proportion of 40%-65%; the solubilizer is a betaine type zwitterionic surfactant, with a weight proportion of 0.5%-3%. The present invention solves the problem of accurately obtaining substantial tissues such as nerves, blood vessels, lymph nodes, cancerous nodules, organ stumps, etc. in adipose tissue in existing surgical gross specimens or biopsy tissue pathological specimens. In subsequent pathological film preparation, all substantial tissues obtained retain their original tissue structures, and do not affect morphological observation, immunohistochemistry, and in situ hybridization effects.

Description

Technical Field

[0001] The invention belongs to the technical field of pathological specimen processing, relates to a pathological specimen processing liquid, and specifically relates to a defatting liquid for permeabilizing surgical gross 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 sampling of surgical 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 has 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 recommended 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 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 in examining tiny solid tissues in the fat tissue of existing surgical gross specimens and the processing defects of current fat dissolving solutions, a new defatting solution for naked eye visualization is provided. The defatting solution has the multiple functions of transparency, fat removal, and fixation of various solid tissues (nerves, blood vessels, lymph nodes, cancer nodules, organ stumps, etc.) in fat tissue. While transparently removing fat tissue, it can effectively retain the morphological structure, immunohistochemical antigen epitopes and in situ hybridization nucleic acid sequences of various solid 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 permeabilization of surgical cadaver specimens, which is composed of four components: a permeabilizing agent, a permeation enhancer, a degreasing agent, and a solubilizing agent. The weight proportions of the four components are as follows: the permeabilizing agent is a five-membered or six-membered azacyclic ketone compound, wherein the N-position substituent is a methyl, ethyl or butyl group, accounting for 2-20% by weight; the permeation enhancer is a C1-C3 lower alcohol, accounting for 30%-55% by weight; the degreasing agent is a C6-C8 cycloalkane, accounting for 40%-65% by weight; and the solubilizing agent is a betaine-type zwitterionic surfactant, accounting for 0.5%-3% 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 0.3-2 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 N-methylpyrrolidone, N-methyl-3-pyrrolidone, N-ethylpyrrolidone, N-ethyl-3-pyrrolidone, N-butylpyrrolidone, caprolactam, and N-methylcaprolactam;

[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 dodecyl dimethyl betaine, dodecyl sulfopropyl betaine, tetradecyl dimethyl betaine, tetradecyl sulfopropyl betaine, octadecyl dimethyl betaine, octadecyl dimethyl sulfopropyl betaine, and cocamidopropyl betaine.

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

[0016] (1) 9.5 parts by weight of N-methylpyrrolidone or N-methyl-3-pyrrolidone, 45 parts by weight of ethanol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of dodecyl dimethyl betaine or dodecyl sulfopropyl betaine;

[0017] (2) 12.5 parts by weight of N-ethylpyrrolidone or N-ethyl-3-pyrrolidone, 40 parts by weight of methanol, 47 parts by weight of methylcyclohexane, and 0.5 parts by weight of tetradecyl dimethyl betaine or tetradecyl sulfopropyl betaine;

[0018] (3) 10 parts by weight of N-butylpyrrolidone, 42 parts by weight of propanol, 47 parts by weight of ethylcyclohexane, and 1 part by weight of octadecyl dimethyl betaine;

[0019] (4) 8.5 parts by weight of caprolactam, 45 parts by weight of methanol, 46 parts by weight of cyclohexane, and 0.5 parts by weight of octadecyldimethylsulfonpropyl betaine;

[0020] (5) 9.5 parts by weight of N-methylcaprolactam, 45 parts by weight of isopropyl alcohol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of cocamidopropyl betaine.

[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 defatting liquid in the transparent treatment of surgical gross specimens or biopsy fat tissue.

[0023] Specifically, the purpose is to defat surgical gross specimens or biopsy tissues to visualize the non-fat tissue inside them with the naked eye. The non-fat tissue includes parenchymal tissues such as nerves, blood vessels, lymph nodes, cancer 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 biopsy tissue is placed in the defatting liquid at a weight-volume ratio of 1:3-1:100 at room temperature, and immersed in the defatting liquid for more than 4 hours, during which the liquid can be changed and immersed repeatedly. 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 surgical gross specimens or biopsy tissues at the same time, and has the functions of transparentizing, degreasing, and fixing specimens. Compared with the formaldehyde treatment liquid commonly used at present, 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 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 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, 5, 9, 10, 11) by naked eye comparison;

[0032] Figure 2 The light transmittance of adipose tissue treated with different degreasing solutions in the control group and the test group (Examples 1 to 11) 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 portal area after cleaning for the control group and the experimental group (Examples 1, 5, 9, 10, 11);

[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, 5, 9, 10, 11);

[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, 5, 9, 10, 11) under microscopic pathology;

[0037] Figure 7 Comparison of paraffin sections and immunohistochemical sections of the organ stump (part of the pancreatic tissue) in the adipose tissue of the liver hilum in the control group and the experimental group (Examples 1, 5, 9, 10, 11) under the microscope;

[0038] Figure 8 Comparison of paraffin sections and immunohistochemical sections of abdominal nodules in the control group and the experimental group (Examples 1, 5, 9, 10, 11) 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 permeabilization of surgical cadaver specimens is shown in Examples 1 to 11:

[0042] Example 1

[0043] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 9.5% N-methylpyrrolidone, 45% ethanol, 45% cyclohexane, and 0.5% dodecyl dimethyl betaine.

[0044] Example 2

[0045] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 9.5% N-methylpyrrolidone, 45% ethanol, 45% cyclohexane, and 0.5% dodecyl sulfopropyl betaine.

[0046] Example 3

[0047] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 9.5% N-methyl-3-pyrrolidone, 45% ethanol, 45% cyclohexane, and 0.5% dodecyl dimethyl betaine.

[0048] Example 4

[0049] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 9.5% N-methyl-3-pyrrolidone, 45% ethanol, 45% cyclohexane, and 0.5% dodecyl sulfopropyl betaine.

[0050] Example 5

[0051] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 12.5% ​​N-ethylpyrrolidone, 40% methanol, 47% methylcyclohexane, and 0.5% tetradecyl dimethyl betaine.

[0052] Example 6

[0053] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 12.5% ​​N-ethylpyrrolidone, 40% methanol, 47% methylcyclohexane, and 0.5% tetradecyl sulfopropyl betaine.

[0054] Example 7

[0055] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 12.5% ​​N-ethyl-3-pyrrolidone, 40% methanol, 47% methylcyclohexane, and 0.5% tetradecyl dimethyl betaine.

[0056] Example 8

[0057] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 12.5% ​​N-ethyl-3-pyrrolidone, 40% methanol, 47% methylcyclohexane, and 0.5% tetradecyl sulfopropyl betaine.

[0058] Example 9

[0059] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 10% N-butylpyrrolidone, 42% propanol, 47% ethylcyclohexane, and 1% octadecyl dimethyl betaine.

[0060] Example 10

[0061] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 8.5% caprolactam, 45% methanol, 46% cyclohexane, and 0.5% octadecyl dimethyl sulfonyl betaine.

[0062] Embodiment 11

[0063] The degreasing solution used for permeabilization of surgical gross specimens in this embodiment includes, based on 100% total weight, 9.5% N-methyl caprolactam, 45% isopropyl alcohol, 45% cyclohexane, and 0.5% cocamidopropyl betaine.

[0064] 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 a degreasing solution for permeabilizing surgical cadaver specimens.

[0065] The above formulas are analyzed and classified according to the permeabilizing agent: Examples 1 to 4 all belong to the N-methylpyrrolidone system; Examples 5 to 8 all belong to the N-ethylpyrrolidone system; Example 9 belongs to the N-butylpyrrolidone system; Example 10 belongs to the caprolactam system; and Example 11 belongs to the caprolactam alkyl substituted system.

[0066] 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 5, Example 9, Example 10 and Example 11 are selected as representatives for subsequent experiments.

[0067] 2. Effect Verification

[0068] 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.

[0069] 1. Experimental materials and reagents

[0070] Experimental materials: Hepatic hilar adipose tissue (surgical specimens) or abdominal nodules (biopsy tissue).

[0071] 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.

[0072] 2. Experimental methods and results

[0073] 2.1 Experimental methods

[0074] 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.

[0075] 2.2 Transparency and transmittance comparison

[0076] The transparency of the fat tissue in the control group and the test group (Examples 1, 5, 9, 10, 11) was observed by naked eyes. The transmittance of the control group and the test group (Examples 1 to 11) was measured using a LS116 transmittance meter. The transmittance measurement method is as follows:

[0077] Transparency measurement of control group:

[0078] (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;

[0079] (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;

[0080] (3) The transmittance (%) was measured using a LS116 transmittance meter. This is the transmittance (%) of the adipose tissue in the control group.

[0081] Determination of transparency of processed tissue:

[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 immersed in the degreasing solution prepared in Examples 1 to 11 at room temperature for 6 hours;

[0084] (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;

[0085] (4) The transmittance (%) was measured using a LS116 transmittance meter. This is the transmittance (%) of the adipose tissue in the experimental group.

[0086] The results of visually comparing the transparency of the fat tissue in the control group and the test group (Examples 1, 5, 9, 10, 11) 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, 5, 9, 10, and 11 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 ).

[0087] Table 1 Comparison of light transmittance between the experimental group and the control group

[0088] Group Light transmittance (%) OP_SD Control group 3.2 0.43 Example 1 31.5 3.26 Example 2 30.2 3.15 Example 3 29.2 2.47 Example 4 28.6 2.39 Example 5 26.7 2.68 Example 6 25.9 3.89 Example 7 27.7 4.36 Example 8 28.4 3.95 Example 9 24.4 3.41 Example 10 29.6 3.35 Embodiment 11 25.3 3.60

[0089] 2.3 Comparison of immunohistochemistry and in situ hybridization

[0090] 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:

[0091] 2.3.1 Morphological observation and immunohistochemical effect of nerve tissue in adipose tissue removed from the hepatic hilum

[0092] The paraffin sections / immunohistochemical sections of the nerve tissue in the hepatic hilar adipose tissue prepared in the test groups (Examples 1, 5, 9, 10, 11) 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.

[0093] 2.3.2 Morphological observation and immunohistochemical effect of vascular tissue in adipose tissue removed from the hepatic hilum

[0094] The paraffin sections / immunohistochemical sections of vascular tissue in the hepatic hilar adipose tissue cleared from the experimental groups (Examples 1, 5, 9, 10, 11) 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.

[0095] 2.3.3 Morphological observation and immunohistochemical effect of lymph nodes in adipose tissue removed from the hepatic hilum

[0096] 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.

[0097] 2.3.4 Morphological observation and immunohistochemical effect of clearing cancer nodules in adipose tissue at the hepatic hilum

[0098] 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, 5, 9, 10, and 11) 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. The results are shown in Figure 6 ,Comparison found that the morphological observation and immunohistochemical effects of the experimental group and the control group were basically the same.

[0099] 2.3.5 Morphological observation and immunohistochemical results of organ stumps in adipose tissue cleared from the hepatic portal (partial pancreatic tissue)

[0100] 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, 5, 9, 10, 11) 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.

[0101] 2.3.6 Morphological observation and immunohistochemical effect on abdominal nodules

[0102] The paraffin sections / immunohistochemical sections of the abdominal nodules prepared in the test groups (Examples 1, 5, 9, 10, and 11) 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.

[0103] 2.3.7 Conclusion

[0104] The degreasing liquid used for permeabilizing surgical gross specimens of the present invention can be used to treat surgical gross specimens or biopsy tissues, and has the functions of making the specimens transparent, degreasing, and fixing the specimens, and does not affect the subsequent morphological observation, immunohistochemistry, and in situ hybridization effects. The liquid is an ideal formula reagent that can be used for pathological specimens of adipose tissues of different volumes.

[0105] 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 permeabilizing surgical cadaver 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 N-methylpyrrolidone, N-methyl-3-pyrrolidone, N-ethylpyrrolidone, N-ethyl-3-pyrrolidone, N-butylpyrrolidone, caprolactam, and N-methylcaprolactam, 2-20 parts by weight; The penetration enhancer is one of methanol, ethanol, propanol and isopropanol, 30-55 parts by weight; The degreasing agent is one of cyclohexane, methylcyclohexane and ethylcyclohexane, 40-65 parts by weight; The solubilizer is one of dodecyl dimethyl betaine, dodecyl sulfopropyl betaine, tetradecyl dimethyl betaine, tetradecyl sulfopropyl betaine, octadecyl dimethyl betaine, octadecyl dimethyl sulfopropyl betaine, and cocamidopropyl betaine, 0.5-3 parts by weight.

2. The degreasing solution for permeabilization of surgical cadaver specimens according to claim 1, characterized in that: in, The weight proportions of the four components are as follows: 5-15 weight parts of a permeabilizing agent, 40-55 weight parts of a penetration enhancer, 45-65 weight parts of a degreasing agent and 0.5-2 weight parts of a solubilizing agent.

3. The degreasing solution for permeabilization of surgical cadaver 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) 9.5 parts by weight of N-methylpyrrolidone or N-methyl-3-pyrrolidone, 45 parts by weight of ethanol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of dodecyl dimethyl betaine or dodecyl sulfopropyl betaine; (2) 12.5 parts by weight of N-ethylpyrrolidone or N-ethyl-3-pyrrolidone, 40 parts by weight of methanol, 47 parts by weight of methylcyclohexane, and 0.5 parts by weight of tetradecyl dimethyl betaine or tetradecyl sulfopropyl betaine; (3) 10 parts by weight of N-butylpyrrolidone, 42 parts by weight of propanol, 47 parts by weight of ethylcyclohexane, and 1 part by weight of octadecyl dimethyl betaine; (4) 8.5 parts by weight of caprolactam, 45 parts by weight of methanol, 46 parts by weight of cyclohexane, and 0.5 parts by weight of octadecyldimethylsulfonpropyl betaine; (5) 9.5 parts by weight of N-methylcaprolactam, 45 parts by weight of isopropyl alcohol, 45 parts by weight of cyclohexane, and 0.5 parts by weight of cocamidopropyl betaine.

4. The method for preparing a defatting solution for permeabilization of surgical cadaver 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 3 in transparentizing surgical gross specimens or biopsy adipose tissue.

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 cadaver specimens or biopsy tissues using the degreasing solution according to any one of claims 1 to 3, characterized in that: Place the surgical specimen or 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 4 hours. During this period, the solution can be changed and the soaking can be repeated.

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