Intestinal content-containing tissue treatment mold and method

Abstract

The invention provides an intestinal tract content-containing tissue treatment mold and method, and relates to the technical field of in vitro detection.The intestinal tract content-containing tissue treatment mold comprises a bottom plate, an intestinal tract model arranged on the bottom plate and a plurality of first dehydration holes formed in the intestinal tract model; the intestinal tract model is of a spiral structure, and the expansion distance of each circle of the spiral structure is 105-110% of the diameter of a sample to be embedded; the cross-sectional area of the first dehydration hole in the direction perpendicular to the axis is 3-6 mm < 2 >, the intestinal tract tissue is kept in a Swiss roll intestine shape after being treated through the intestinal tract model, and adhesion of adjacent intestinal tract walls is avoided. The intestinal tissue is better embedded into the spiral structure and is in full contact with a solution, and the problems that in the prior art, after the intestinal tissue containing the content is dehydrated, the complete Swiss roll shape cannot be kept and slices are incomplete under the condition that the tissue is not damaged, or the curled intestinal tissue walls are prone to adhesion, and the intestinal tissue is damaged are solved. And in-situ capture of the space-time transcriptome is influenced after slicing, and the space-time transcriptome is restored to a spatial

Description

Technical Field

[0001] The present invention relates to the technical field of in vitro detection, and in particular to a mold and a method for processing intestinal content tissue. Background Art

[0002] Current epidemiological, pathological, omics, cell and animal research results reveal that intestinal microorganisms mediate people's metabolic health and disease risks to a considerable extent. The collection of these tiny organisms that parasitize various cavities in the human body is called microorganisms, including a large number of bacteria, fungi, bacteriophages, eukaryotic viruses and fungi. Most of them are symbiotic or mutualistic microorganisms. The intestinal microbiome plays an important role in improving host immunity, food digestion, intestinal endocrine function and neural signal regulation, drug function and metabolism, endotoxin clearance and affecting the production of host metabolism-related substances. In the past two decades, the conclusions from observational studies have naturally led people to think that intestinal flora may affect the metabolic homeostasis of the host, and disordered intestinal flora can lead to the occurrence of many common metabolic diseases, including obesity, type 2 diabetes, non-alcoholic liver disease, metabolic heart disease and malnutrition. In order to obtain a mechanistic understanding of the influence of intestinal flora on host metabolism, the research perspective has shifted from the traditional descriptive analysis of flora composition to the exploration of causal relationships. In addition to the reported correlation results, the integration of high-throughput multi-omics data from various types of populations, such as metagenomics, metabolomics, etc., as well as host physiological phenotypic characteristics and experimental results from clinical, animal, and cell models, is playing an increasingly important and supporting role in clarifying the role of intestinal flora in human metabolic diseases.

[0003] Comparative analysis of fecal samples and intestinal flora in different intestinal segments found that fecal samples had the highest similarity with the intestinal flora of the colon. However, no spatial omics study of the entire intestinal tract has been conducted in current research.

[0004] With the promotion of spatiotemporal transcriptome FFPE (Formalin-Fixed Paraffin Embedded) product solutions, the TotalRNA molecules in tissue cells in FFPE samples are captured in situ using "random probes" and restored to spatial positions through spatial barcodes (CoordinateID, CID), which can realize the construction of spatial expression maps of the entire transcriptome of all species. Compared with common transcriptome solutions, this solution can capture degraded RNA fragments, and can also achieve co-capture of coding and non-coding RNA, as well as co-detection of hosts and microorganisms, bringing new opportunities for scientific research. However, in the preparation of traditional paraffin-embedded samples, tissue sections containing contents in the intestine have not been made. On the one hand, after paraffin embedding, the RNA in the intestine degrades, affecting extraction and research analysis; on the other hand, after the intestinal contents are dehydrated by conventional dehydration procedures, the contents become hard and the tissue cannot be cut out of a complete shape. Summary of the invention

[0005] One of the purposes of the present invention is to provide an intestinal tissue processing mold containing contents, so as to at least solve the technical problems in the prior art that the intestinal tissue containing contents cannot maintain a complete Swiss roll shape after dehydration treatment, resulting in incomplete slicing, or that adhesion is easily caused between the curled intestinal tissue walls, which affects the in situ capture of the spatiotemporal transcriptome after slicing and restoration to the spatial position.

[0006] A second object of the present invention is to provide a method for processing intestinal tissue containing contents.

[0007] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0008] In a first aspect, the present invention provides an intestinal tissue processing mold containing contents, comprising a bottom plate, an intestinal model arranged on the bottom plate, and a plurality of first dehydration holes arranged on the intestinal model;

[0009] The intestinal model is a spiral structure, and the extension distance of each circle of the spiral structure is 105-110% of the diameter of the sample to be embedded;

[0010] The cross-sectional area of ​​the first dehydration hole in the vertical axis direction is 3 to 6 mm 2 .

[0011] Furthermore, the bottom plate is provided with a plurality of second dehydration holes, and the cross-sectional area of ​​the second dehydration holes in the direction perpendicular to the axis is larger than the cross-sectional area of ​​the first dehydration holes in the direction perpendicular to the axis.

[0012] Furthermore, the cross-sectional area of ​​the second dehydration hole in the vertical axis direction is 7 to 12 mm 2 , preferably 10mm 2;

[0013] Preferably, the height of the intestinal model is 3.5 to 10 mm;

[0014] Preferably, the thickness of the intestinal model is 0.5-1 mm.

[0015] Furthermore, the base plate and the intestinal model are an integrated structure.

[0016] In a second aspect, the present invention provides a method for processing intestinal tissue containing contents, comprising placing the intestinal tissue containing contents in the above-mentioned intestinal tissue containing contents processing mold, performing fixation treatment and dehydration treatment in sequence, and then taking out the intestinal tissue.

[0017] Furthermore, the placement method includes embedding the intestinal tissue containing the contents in the spiral structure.

[0018] Furthermore, before the dehydration treatment, the method further includes placing the intestinal content-containing tissue processing mold into an embedding box;

[0019] Preferably, the dehydration treatment comprises sequentially immersing in solutions of the following concentrations for dehydration:

[0020]

[0021]

[0022] Furthermore, the intestinal tract includes any one of the duodenum, jejunum, ileum, colon or rectum.

[0023] Furthermore, when the intestinal tissue is ileum tissue or colon tissue, the soaking time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration treatment is 30 to 40 minutes;

[0024] Preferably, when the intestinal tissue is rectal tissue, the immersion time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration treatment is 30 minutes.

[0025] Furthermore, the fixation treatment includes soaking in a fixative solution for 24 to 72 hours;

[0026] Preferably, the fixative comprises a tissue fixative containing 10% DEPC, paraformaldehyde or NBF with a mass concentration of 10%.

[0027] The present invention provides a mold for processing intestinal tissue containing contents. Through the intestinal model, the intestinal tissue maintains the Swiss roll shape after the fixation and dehydration steps to achieve the fixation effect. The intestinal model can also avoid the adhesion of adjacent intestinal walls to ensure that the spatial position relationship of the contents is not destroyed. By limiting the extension distance of each circle of the spiral structure, the intestinal tissue can be better embedded in the spiral structure, and the intestinal tissue contents are prevented from hardening or shrinking during the fixation and dehydration process, so that the intestinal tissue is loosened and falls off the mold or cannot be completely removed after treatment. At the same time, the solution in the fixation and dehydration process can penetrate into the gap between the intestinal tissue and the spiral structure to wrap the intestinal tissue. At the same time, the first dehydration hole is set to enable the intestinal tissue to fully contact with the fixative and the solutions in each stage of the dehydration process, reduce the influence of the mold blocking the contact between the solution and the intestinal tissue in the fixation and dehydration stages, and improve the use effect of the mold.

[0028] On the other hand, a method for processing intestinal tissue containing contents is provided, which reduces the difficulty of operation during the fixation process and maintains the integrity of the tissue morphology by using auxiliary tools.

[0029] The present invention solves the technical problems in the prior art that the intestinal tissue containing contents cannot maintain a complete Swiss roll shape after dehydration treatment, resulting in incomplete slicing, or that the curled intestinal tissue walls are prone to adhesion, which affects the in situ capture of the spatiotemporal transcriptome after slicing and the restoration to the spatial position. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0031] Figure 1 A schematic diagram of the three-dimensional structure of a mold for processing intestinal tissue containing contents provided in Example 1 of the present invention;

[0032] Figure 2 A schematic diagram of the top view of a mold for processing intestinal tissue containing contents provided in Example 1 of the present invention;

[0033] Figure 3 A schematic side view of the structure of a mold for processing intestinal tissue containing contents provided in Example 1 of the present invention;

[0034] Figure 4 A comparison chart of the treatment effects of different intestinal content tissues provided in Example 5 of the present invention;

[0035] Figure 5 A comparison diagram of the treatment effects of intestinal content tissue provided in Example 9 of the present invention;

[0036] Figure 6 A comparison diagram of the treatment effects of intestinal content tissue provided in Example 10 of the present invention;

[0037] Figure 7 A comparison chart of the treatment effects of different molds provided by the present invention on intestinal content tissue;

[0038] Figure 8 This is a diagram showing the effect of treating the intestinal contents provided in Comparative Example 7 of the present invention;

[0039] Fig. 9 This is a diagram showing the effect of intestinal content tissue processing provided in Comparative Example 8 of the present invention.

[0040] Icon: 1- bottom plate; 2- intestinal model; 3- first dehydration hole; 4- second dehydration hole. DETAILED DESCRIPTION

[0041] Unless otherwise defined herein, scientific and technical terms used in conjunction with the present invention shall have the meanings commonly understood by those of ordinary skill in the art. The meaning and scope of the terms should be clear, however, in the case of any potential ambiguity, the definitions provided herein take precedence over any dictionary or external definitions. In this application, unless otherwise stated, the use of "or" means "and / or". In addition, the use of the term "including" and other forms is non-limiting.

[0042] The methods and techniques of the present invention are generally performed according to conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification unless otherwise indicated.

[0043] On one hand, the present invention provides an intestinal tissue processing mold containing contents, comprising a bottom plate 1, an intestinal model 2 arranged on the bottom plate 1, and a plurality of first dehydration holes 3 arranged on the intestinal model 2;

[0044] The intestinal model 2 is a spiral structure, and the extension distance of each circle of the spiral structure is 105-110% of the diameter of the sample to be embedded;

[0045] The cross-sectional area of ​​the first dehydration hole 3 in the vertical axis direction is 3-6 mm 2 .

[0046] Through the intestinal model 2, the intestinal tissue maintains the Swiss roll shape after the fixation and dehydration steps to achieve the fixation effect, and the intestinal model 2 can also avoid the adhesion of adjacent intestinal walls to ensure that the spatial position relationship of the contents is not destroyed. By limiting the extension distance of each circle of the spiral structure, the intestinal tissue can be better embedded in the spiral structure, and the contents of the intestinal tissue are prevented from hardening or shrinking during the fixation and dehydration process, so that the intestinal tissue loosens and falls off from the mold or cannot be completely removed after treatment. At the same time, the solution in the fixation and dehydration process can penetrate from the gap between the intestinal tissue and the spiral structure to wrap the intestinal tissue. At the same time, the first dehydration hole 3 is set to enable the intestinal tissue to fully contact with the fixative and the solution of each stage of the dehydration process, reduce the influence of the mold blocking the contact between the solution and the intestinal tissue in the fixation and dehydration stages, and improve the use effect of the mold. It solves the technical problems that the intestinal tissue containing contents cannot maintain a complete Swiss roll shape after dehydration treatment, resulting in incomplete slicing, or the curled intestinal tissue walls are prone to adhesion, which affects the in situ capture of the spatiotemporal transcriptome after slicing and restores it to the spatial position.

[0047] The expansion distance of each circle refers to the distance between the outer circle and the inner adjacent circle in the direction of outward expansion of the spiral model.

[0048] In some specific embodiments, a plurality of second dehydration holes 4 are further provided on the bottom plate 1 , and the cross-sectional area of ​​the second dehydration holes 4 in the direction perpendicular to the axis is larger than the cross-sectional area of ​​the first dehydration holes 3 in the direction perpendicular to the axis.

[0049] In some specific embodiments, the cross-sectional area of ​​the second dehydration hole 4 in the direction perpendicular to the axis is 7 to 12 mm. 2 .

[0050] The cross-sectional area of ​​the second dehydration hole 4 in the vertical axis direction can be, but is not limited to, 7 mm 2 , 8mm 2 , 9mm 2 , 10mm 2 、11mm 2 or 12mm 2 , can also be 7 to 12 mm 2 Any value between, preferably 10mm 2 .

[0051] In some specific embodiments, the height of the intestinal model 2 is 3.5 to 10 mm.

[0052] Among them, the height of the intestinal model 2 can be but is not limited to 3.5mm, 4mm, 4.5mm, 5mm, 5.5mm, 6mm, 6.5mm, 7mm, 7.5mm, 8mm, 8.5mm, 9mm, 9.5mm or 10mm, and can also be any value between 3.5 and 10mm.

[0053] In some specific embodiments, the thickness of the intestinal model 2 is 0.5 to 1 mm.

[0054] The thickness of the intestinal model 2 may be, but is not limited to, 0.5 mm, 0.6 mm, 0.7 mm, 0.8 mm, 0.9 mm or 1 mm, and may be any value between 0.5 and 1 mm, preferably 0.8 mm.

[0055] In some specific embodiments, the base plate 1 and the intestinal model 2 are an integrated structure.

[0056] According to another aspect of the present invention, a method for processing intestinal tissue containing contents is provided, comprising placing the intestinal tissue containing contents in the above-mentioned intestinal tissue containing contents processing mold, performing fixation and dehydration treatment in sequence, and then removing the intestinal tissue. By using auxiliary tools, the difficulty of operation during the fixation process is reduced, and the integrity of the tissue morphology is maintained.

[0057] In some specific embodiments, the method of placing includes embedding the intestinal tissue containing the contents in the spiral structure, wherein embedding means that the intestinal tissue is placed in the spiral structure without any excess intestinal tissue protruding outside the spiral structure.

[0058] In some specific embodiments, the dehydration treatment further includes placing the intestinal content tissue processing mold into an embedding box before the dehydration treatment. Based on this model, the dehydration procedure is optimized by using the tissue embedding box in the dehydration process. In some specific embodiments, the dehydration treatment includes sequentially immersing in the following concentrations of solution for dehydration:

[0059]

[0060] By controlling the soaking time in anhydrous ethanol, the dehydration effect is improved so that each intestinal segment containing the contents can be cut into a complete shape.

[0061] Among them, the immersion time of anhydrous ethanol 1 and anhydrous ethanol 2 can be independently but not limited to 20min, 22min, 24min, 26min, 28min, 30min, 32min, 34min, 36min, 38min or 40min, and can also be any value between 20 and 40min.

[0062] The numbers in 95% ethanol 1, 95% ethanol 2, anhydrous ethanol 1, anhydrous ethanol 2, xylene 1, xylene 2, paraffin 1, paraffin 2 and paraffin 3 only represent the order of immersion in the same reagent.

[0063] In some specific embodiments, the intestinal tract includes any one of the duodenum, jejunum, ileum, colon or rectum.

[0064] Experiments have shown that the change of the immersion time of anhydrous ethanol has different treatment effects on intestinal tissues from different intestinal segments. In some specific embodiments, when the intestinal tissue is ileum tissue or colon tissue, the immersion time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration treatment is 30 to 40 minutes. In some specific embodiments, when the intestinal tissue is rectal tissue, the immersion time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration treatment is 30 minutes. Different immersion times can be selected according to the specific type of intestinal tissue to improve the treatment effect and ensure the integrity of the slices after embedding.

[0065] In some specific embodiments, the fixation treatment includes immersing in a fixative for 24 to 72 hours. Specifically, the immersion time may be 24 hours, 34 hours, 44 hours, 54 hours, 64 hours or 72 hours, or any value between 24 and 72 hours.

[0066] In some specific embodiments, the fixing solution includes paraformaldehyde or NBF with a mass concentration of 10%.

[0067] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0068] The 3D printing equipment was Aurora Elvo Z-603S, and the printing material was polyoxymethylene (POM), which was purchased from Aurora Elvo Technology Co., Ltd.

[0069] Example 1

[0070] Combination Figure 1 to Figure 3 To illustrate, a mold for processing intestinal tissue containing contents includes a base plate 1, an intestinal model 2 arranged on the base plate 1, and a first dehydration hole 3 arranged on the intestinal model 2. Several first dehydration holes 3 are arranged in sequence along the extension direction of the intestinal model 2, so that the part of the intestinal model 2 used for embedding intestinal tissue is arranged with the first dehydration holes 3.

[0071] The intestinal model 2 is a spiral structure, and the extension distance of each circle of the spiral structure is 105% of the diameter of the sample to be embedded; the cross-sectional area of ​​the first dehydration hole 3 in the vertical axis direction is 4mm 2 The cross-sectional area of ​​the second dehydration hole 4 on the bottom plate 1 in the vertical axis direction is 10 mm 2 The height of the intestinal model 2 is 4 mm and the thickness is 0.8 mm.

[0072] By using 3D printing technology, a spiral model that meets the requirements is produced, and the spiral structures of the base plate 1 and the intestinal model 2 are an integrated structure.

[0073] Example 2

[0074] Different from Example 1, the intestinal model 2 is a spiral structure, and the extension distance of each circle of the spiral structure is 108% of the diameter of the sample to be embedded; the cross-sectional area of ​​the first dehydration hole 3 in the vertical axis direction is 6mm 2 The cross-sectional area of ​​the second dehydration hole 4 on the bottom plate 1 in the vertical axis direction is 12 mm 2 The intestinal model 2 has a height of 4.5 mm and a thickness of 1 mm.

[0075] Example 3

[0076] Different from Example 1, the intestinal model 2 is a spiral structure, and the extension distance of each circle of the spiral structure is 110% of the diameter of the sample to be embedded; the cross-sectional area of ​​the first dehydration hole 3 in the vertical axis direction is 3mm 2 The cross-sectional area of ​​the second dehydration hole 4 on the bottom plate 1 in the vertical axis direction is 7mm 2 The height of the intestinal model 2 is 3.5 mm and the thickness is 0.5 mm.

[0077] Example 4

[0078] The difference from Example 1 is that the cross-sectional area of ​​the second dehydration hole 4 in the vertical axis direction is 4 mm 2 .

[0079] Comparative Example 1

[0080] Different from Example 1, the first dehydration hole 3 and the second dehydration hole 4 are not provided on the intestinal model 2.

[0081] Comparative Example 2

[0082] Different from Example 1, the cross-sectional area of ​​the first dehydration hole 3 on the intestinal model 2 in the vertical axis direction is 30 mm 2 .

[0083] Comparative Example 3

[0084] Different from Example 1, the intestinal model 2 is a spiral structure, and the extension distance of each circle of the spiral structure is 150% of the diameter of the sample to be embedded.

[0085] Example 5

[0086] A method for processing intestinal tissue containing contents, wherein the intestinal tissue sample containing contents includes duodenum tissue, jejunum tissue, ileum tissue, colon tissue and rectum tissue. After the intestinal tissue is removed from the body, the excess fat tissue on the intestinal tract is trimmed and the tissue of the corresponding intestinal segment is cut. The diameter of the small intestine is about 3.0 mm, and the diameter of the large intestine is about 3.5 mm. The intestinal tissue samples are processed according to the following steps:

[0087] Take the molds provided in Example 1 and embed the intestinal tissue exactly in the middle of the intestinal model 2. Place the intestinal tissue on the model and fix it in the fixative for 24 hours. Put the fixed tissue together with the fixed mold into the finished embedding box for dehydration.

[0088] The program set for the automatic dehydrator is: tissue fixative containing 10% DEPC for 2 hours; 75% ethanol for 1 hour; 80% ethanol for 1 hour; 95% ethanol for 1 hour; 95% ethanol for 2 hours; anhydrous ethanol for 30 minutes; anhydrous ethanol for 30 minutes; xylene for 30 minutes; xylene for 30 minutes; paraffin for 1 hour; paraffin for 1 hour; paraffin for 2 hours; take out the dehydrated tissue, embed it in paraffin, make paraffin-embedded blocks, perform paraffin sectioning, HE staining, and obtain the sectioning results.

[0089] like Figure 4 The a to e shown are duodenum tissue, jejunum tissue, ileum tissue, colon tissue and rectum tissue respectively. The intestinal tissue fixed by the mold has a complete structure and is in the shape of a Swiss roll sausage. After dehydration, it does not deform. The mold is used in conjunction with the dehydration process. All the intestinal slices containing the contents are intact and do not affect the histopathological observation and judgment.

[0090] Embodiments 6 to 8

[0091] The difference from Example 5 is that the molds provided in Examples 2 to 4 are used for the operation, and the intestinal tissue sample containing the contents is the ileum. Figure 7 As shown, d to f are Examples 2 to 4 respectively. It can be seen that in Examples 2 and 3, the intestinal tissues are not deformed and the cut slices are intact. In Example 4, since the cross-sectional area of ​​the second dehydration hole 4 in the direction perpendicular to the axis is equal to the cross-sectional area of ​​the first dehydration hole 3 in the direction perpendicular to the axis, the contact between the solution and the intestinal tissue during the fixation and dehydration treatments is affected, and the intestinal tissue is slightly deformed, but the cut slices are intact, which does not affect the histopathological observation and judgment.

[0092] Example 9

[0093] The difference from Example 5 is that the tissue fixed with the mold is dehydrated, and the soaking time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration procedure is shortened to 40 minutes. After dehydration, the tissue is embedded, sliced ​​and stained with HE to obtain Figure 5 Results shown.

[0094] like Figure 5 As shown, a to d are duodenum tissue, jejunum tissue, ileum tissue and colon tissue, respectively. The duodenum, jejunum, ileum and colon can all be cut out in complete shape, indicating that adjusting the immersion time of anhydrous ethanol is effective, but the rectum has a lot of hard contents and cannot be cut into complete sections.

[0095] Example 10

[0096] The difference from Example 5 is that the tissue fixed with the mold is dehydrated, and the soaking time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration procedure is shortened to 20 min. After dehydration, the tissue is embedded, sliced ​​and HE stained to obtain Figure 6 The results are shown, where a to d are duodenum, jejunum, colon and rectum, respectively. In this dehydration procedure, the ileum, which contains rich intestinal contents, was not cut out, and the duodenum, jejunum, colon and rectum were all cut out, but the colon and rectum were not cut into sections with complete morphology.

[0097] Comparative Examples 4 to 6

[0098] The difference from Example 5 is that the molds provided in Comparative Examples 1 to 3 were used to process the intestinal tissue samples (ileum) containing the contents. After dehydration, tissue embedding, tissue sectioning and HE staining were performed to obtain Figure 7 The results are shown, where a to c are comparative examples 1 to 3 respectively. As shown in the HE staining results, compared with Example 5, in comparative example 1, since the first dehydration hole 3 and the second dehydration hole 4 are not provided, the contact area between the tissue and the fixative and the dehydration reagent is reduced, so the tissue section results show that the tissue is wrinkled and broken, which affects the section quality and pathological section observation; in comparative example 2, the length of the first dehydration hole 3 is increased, and the tissue is deformed during the fixation and dehydration process, and the tissue cannot be embedded in the same cutting surface, which affects the embedding effect; in comparative example 3, the diameter of the expansion distance of each circle of the spiral structure is increased, and the tissue is deformed during the fixation and dehydration process, and the tissue cannot be embedded in the same cutting surface, which affects the embedding effect.

[0099] Comparative Example 7

[0100] Different from Example 5, in the fixation process, the jejunal tissue was directly fixed without using a mold, and the slice results were obtained after fixation, dehydration, embedding, and staining. Figure 8As shown in the figure, during the fixation process, the shape of the intestinal tissue is not fixed, so after fixation and dehydration, during the embedding process, the tissue cannot be completely embedded due to the limited size of the embedding mold. After staining, the staining result is obtained. Due to the incomplete tissue and the breakage during the embedding process, the tissue is artificially damaged, which increases the difficulty of pathological reading.

[0101] Comparative Example 8

[0102] The difference from Example 5 is that the mold of Example 1 is used for fixation during the fixation process. Before the dehydration process, the tissue is removed from the mold and directly placed in an embedding box for dehydration, followed by tissue embedding and HE staining. Fig. 9 As shown, the morphology of the tissue did not change during the fixation process, but during the dehydration process, due to the lack of mold restrictions, tissue dehydration shrinkage and the external force of the reagent, the Swiss roll sausage structure became loose. During the embedding process, the intestinal tissue could not be placed on the same embedding surface, affecting the integrity of the tissue section.

[0103] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A mold for processing intestinal contents tissue, characterized in that: It comprises a bottom plate (1), an intestinal model (2) arranged on the bottom plate (1), and a plurality of first dehydration holes (3) arranged on the intestinal model (2); The intestinal model (2) is a spiral structure, and the extension distance of each turn of the spiral structure is 105-110% of the diameter of the sample to be embedded; The cross-sectional area of ​​the first dehydration hole (3) in the direction perpendicular to the axis is 3 to 6 mm 2 .

2. The intestinal content tissue processing mold according to claim 1, characterized in that: The bottom plate (1) is also provided with a plurality of second dehydration holes (4), and the cross-sectional area of ​​the second dehydration holes (4) in the direction perpendicular to the axis is larger than the cross-sectional area of ​​the first dehydration holes (3) in the direction perpendicular to the axis.

3. The intestinal content tissue processing mold according to claim 2, characterized in that: The cross-sectional area of ​​the second dehydration hole (4) in the direction perpendicular to the axis is 7 to 12 mm 2 , preferably 10mm 2 ; Preferably, the height of the intestinal model (2) is 3.5 to 10 mm; Preferably, the thickness of the intestinal model (2) is 0.5-1 mm.

4. The intestinal content tissue processing mold according to any one of claims 1 to 3, characterized in that: The base plate (1) and the intestinal model (2) are an integrated structure.

5. A method for processing intestinal tissue containing contents, characterized in that: The method comprises placing the intestinal tissue containing contents in the intestinal tissue containing contents processing mold according to any one of claims 1 to 4, performing fixation treatment and dehydration treatment in sequence, and then taking out the intestinal tissue.

6. The method for processing intestinal tissue containing contents according to claim 5, characterized in that: The method of placement includes embedding the intestinal tissue containing the contents in a spiral structure.

7. The method for processing intestinal tissue containing contents according to claim 5, characterized in that: Before the dehydration treatment, the intestinal tissue processing mold containing the intestinal contents is placed in an embedding box; Preferably, the dehydration treatment comprises sequentially immersing in solutions of the following concentrations for dehydration:

8. The method for processing intestinal tissue containing contents according to claim 7, characterized in that: The intestinal tract includes any one of the duodenum, jejunum, ileum, colon or rectum.

9. The method for processing intestinal tissue containing contents according to claim 8, characterized in that: When the intestinal tissue is ileum tissue or colon tissue, the soaking time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration treatment is 30 to 40 minutes; Preferably, when the intestinal tissue is rectal tissue, the immersion time of anhydrous ethanol 1 and anhydrous ethanol 2 in the dehydration treatment is 30 minutes.

10. The method for processing intestinal tissue containing contents according to any one of claims 5 to 9, characterized in that: The fixation treatment comprises soaking in a fixative for 24 to 72 hours; Preferably, the fixative comprises a tissue fixative containing 10% DEPC, paraformaldehyde or NBF with a mass concentration of 10%.

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