Intestinal content-containing tissue treatment mold
By designing a spiral intestinal model and dehydration pore structure, the problems of intestinal tissue incomplete and adhesion after dehydration treatment are solved, ensuring section integrity and accurate reduction of the spatiotemporal transcriptome.
Patent Information
- Application Number
- CN202422739708.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-11
- Publication Date
- 2025-09-05
- Estimated Expiration
- 2034-11-11
AI Technical Summary
In the prior art, the intestinal content-containing tissue cannot maintain the complete Swiss roll shape after dehydration treatment, resulting in incomplete sections or prone to adhesions between intestinal tissue walls, affecting the effect of in situ capture of the space-time transcriptome.
The intestinal model with a spiral structure and the first dehydration hole arranged on it ensure that the intestinal tissue remains intact during the fixation and dehydration process and avoids adhesion. The intestinal tissue is embedded and fully in contact with the solution through the design of the spiral structure, which solves the deformation and adhesion problems of intestinal tissue during the treatment process.
The morphology of intestinal tissue during fixation and dehydration is achieved, ensuring section integrity, supporting in situ capture of the spatiotemporal transcriptome and accurate reduction of spatial location.
Smart Images
Figure CN223307956U_ABST
Abstract
Description
Technical Field
[0001] The utility model relates to the technical field of embedding, in particular to a mold for processing intestinal tissue containing contents. Background Art
[0002] Current epidemiological, pathological, omics, cell-based, and animal studies reveal that the intestinal microbiome significantly influences human metabolic health and disease risk. The microbiome, a collection of tiny organisms that inhabit various cavities of the human body, encompasses a vast array of bacteria, fungi, bacteriophages, eukaryotic viruses, and fungi. The majority of these microorganisms are commensal or mutualistic. The gut microbiome plays a crucial role in enhancing host immunity, food digestion, intestinal endocrine function and neural signaling regulation, drug function and metabolism, endotoxin clearance, and the production of substances that influence host metabolism. Over the past two decades, findings from observational studies have led to the assumption that the gut microbiome may influence host metabolic homeostasis. Disturbances in the gut microbiome contribute to the development of numerous common metabolic diseases, including obesity, type 2 diabetes, non-alcoholic liver disease, metabolic heart disease, and malnutrition. To gain a mechanistic understanding of how the gut microbiome influences host metabolism, research has shifted from traditional descriptive analyses of microbial composition to investigations 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 research has yet conducted spatial omics studies on the entire intestinal tract.
[0004] With the widespread adoption of spatiotemporal transcriptomics FFPE (Formalin-Fixed Paraffin Embedded) product solutions, total RNA molecules within tissue cells in FFPE samples can be captured in situ using "random probes" and restored to their spatial positions using spatial barcodes (Coordinate ID, CID), enabling the construction of spatial expression maps of the entire transcriptome across all species. Compared to common transcriptomics solutions, this solution can capture degraded RNA fragments, co-capture coding and non-coding RNAs, and co-detect host and microbial activity, bringing new opportunities to scientific research. However, traditional paraffin-embedded sample preparation has not yet involved tissue sections containing intestinal contents. On the one hand, RNA in the intestine degrades after paraffin embedding, hindering extraction and research analysis. On the other hand, after conventional dehydration procedures, the intestinal contents become hardened, making it difficult to slice the tissue in its entirety. Utility Model Content
[0005] The purpose of the present utility model is to provide a mold for processing intestinal tissue 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 easily occurs 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] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are adopted:
[0007] The utility model provides an intestinal tissue processing mold containing contents, which is characterized by comprising a bottom plate, an intestinal model arranged on the bottom plate, and a plurality of first dehydration holes arranged on the intestinal model;
[0008] The intestinal model 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;
[0009] The cross-sectional area of the first dehydration hole in the vertical axis direction is 3 to 6 mm 2 .
[0010] Furthermore, a plurality of second dehydration holes (4) are provided on the bottom plate (1).
[0011] Furthermore, the cross-sectional area of the second dehydration hole (4) in the direction perpendicular to the axis is greater than the cross-sectional area of the first dehydration hole (3) 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 .
[0013] Furthermore, the cross-sectional area of the first dehydration hole in the vertical axis direction is 4mm 2 The cross-sectional area of the second dehydration hole in the vertical axis direction is 10mm 2 .
[0014] Furthermore, the height of the intestinal model is 3.5 to 10 mm.
[0015] Furthermore, the height of the intestinal model is 4 mm.
[0016] Furthermore, the thickness of the intestinal model is 0.5 to 1 mm.
[0017] Furthermore, the thickness of the intestinal model is 0.8 mm.
[0018] Furthermore, the base plate and the intestinal model are an integrated structure.
[0019] The present invention provides a mold for processing intestinal tissue containing contents. The mold maintains the intestinal tissue in a Swiss roll shape after fixation and dehydration, achieving a fixed effect. The mold also prevents adhesion between adjacent intestinal walls, ensuring that the spatial relationship of the contents is not disrupted. By limiting the extension distance of each turn of the spiral structure, the intestinal tissue can be better embedded in the spiral structure, preventing the contents of the intestinal tissue from hardening or shrinking during fixation and dehydration, causing the intestinal tissue to loosen and fall off the mold or become incomplete after treatment. Furthermore, the mold allows the solution during fixation and dehydration to penetrate through the gap between the intestinal tissue and the spiral structure to wrap the intestinal tissue. The provision of a first dehydration hole allows the intestinal tissue to fully contact the fixative and solutions during each dehydration stage, reducing the impact of the mold blocking the contact between the solution and the intestinal tissue during the fixation and dehydration stages and improving the mold's performance. The mold solves the existing technical problems of intestinal tissue containing contents failing to maintain a complete Swiss roll shape after dehydration, resulting in incomplete slicing, or adhesion between the curled intestinal tissue walls, which affects the in situ capture of the spatiotemporal transcriptome and restoration of its spatial position after slicing. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] In order to more clearly illustrate the specific implementation methods of the utility model 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 utility model. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.
[0021] Figure 1A schematic diagram of the three-dimensional structure of a mold for processing intestinal tissue containing contents provided in Example 1 of the present utility model;
[0022] Figure 2 This is a schematic top view of a mold for processing intestinal tissue containing contents provided in Example 1 of the present utility model;
[0023] Figure 3 This is a side structural schematic diagram of a mold for processing intestinal tissue containing contents provided in Example 1 of the present utility model;
[0024] Figure 4 This is a comparison chart of the effects of the mold provided in Example 1 of the present utility model on treating different intestinal tissues containing contents;
[0025] Figure 5 This is a comparison chart of the effects of using different molds to treat intestinal tissue containing contents provided in Experiment 2 of the present invention;
[0026] Figure 6 This is a diagram showing the effect of processing intestinal tissue containing contents without using a mold, provided in Experiment 3 of the present invention.
[0027] Icon: 1-base plate; 2-intestinal model; 3-first dehydration hole; 4-second dehydration hole. DETAILED DESCRIPTION
[0028] Unless otherwise defined herein, scientific and technical terms used in conjunction with this utility model 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 event 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.
[0029] In the description of this utility model, it should be noted that the terms "proximal end," "distal end," "front end," "rear end," "inner," and "outer," etc., indicating positions or relationships, are based on the positions or relationships shown in the accompanying drawings, or are the positions or relationships in which the utility model product is typically placed when in use. These terms are intended solely to facilitate the description of this utility model and simplify the description, and are not intended to indicate or imply that the device or component referred to must have a specific orientation, be constructed, or operate in a specific orientation. Therefore, they should not be construed as limitations on this utility model. Furthermore, the terms "first," "second," etc., etc., are used solely for distinction and should not be construed as indicating or implying relative importance.
[0030] It should also be noted that, in the description of this utility model, unless otherwise expressly specified or limited, the terms "disposed," "installed," and "connected" should be understood broadly. For example, they may refer to fixed connections, detachable connections, or integral connections; they may refer to direct connections, indirect connections through an intermediate medium, or internal connections between two components. Those skilled in the art will understand the specific meanings of these terms in this utility model based on the specific circumstances.
[0031] The utility model provides an intestinal tissue processing mold containing intestinal contents, comprising a base plate 1, an intestinal model 2 arranged on the base plate 1, and a plurality of first dehydration holes 3 arranged on the intestinal model 2;
[0032] 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;
[0033] The cross-sectional area of the first dehydration hole 3 in the vertical axis direction is 3 to 6 mm. 2 .
[0034] The intestinal model 2 allows the intestinal tissue to maintain the Swiss roll shape after the fixation and dehydration steps to achieve a fixation effect, and the intestinal model 2 can also prevent the adjacent intestinal walls from adhering to each other 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, preventing the contents of the intestinal tissue from hardening or shrinking during the fixation and dehydration process, causing the intestinal tissue to loosen and fall off the mold or be unable to be completely removed after treatment. At the same time, the solution during 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 3 is provided to enable the intestinal tissue to fully contact with the fixative and the solution of each stage of the dehydration process, reducing the impact of the mold blocking the contact between the solution and the intestinal tissue during the fixation and dehydration stages, and improving the use effect of the mold. It 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 restores it to the spatial position.
[0035] The expansion distance of each circle refers to the distance between the outer circle and the adjacent inner circle of the spiral model along the outward expansion direction.
[0036] In some specific embodiments, the bottom plate 1 is further provided with a plurality of second dehydration holes 4 to facilitate the mold to be immersed in the solution.
[0037] In some specific embodiments, the cross-sectional area of the second dehydration hole 4 in the direction perpendicular to the axis is larger than the cross-sectional area of the first dehydration hole 3 in the direction perpendicular to the axis.
[0038] In some specific embodiments, the cross-sectional area of the second dehydration hole 4 in the vertical axis direction is 7 to 12 mm. 2 .
[0039] 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 In some specific embodiments, the cross-sectional area of the second dehydration hole 4 in the vertical axis direction is 10 mm 2 .
[0040] In some specific embodiments, the height of the intestinal model 2 is 3.5 to 10 mm.
[0041] The height of the intestinal model 2 may be, but is not limited to, 3.5 mm, 4 mm, 4.5 mm, 5 mm, 5.5 mm, 6 mm, 6.5 mm, 7 mm, 7.5 mm, 8 mm, 8.5 mm, 9 mm, 9.5 mm, or 10 mm, or any value between 3.5 and 10 mm, preferably 4 mm. In some specific embodiments, the height of the intestinal model 2 is 4 mm.
[0042] In some specific embodiments, the thickness of the intestinal model 2 is 0.5 to 1 mm.
[0043] 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, or any value between 0.5 and 1 mm, preferably 0.8 mm. In some specific embodiments, the thickness of the intestinal model 2 is 0.8 mm.
[0044] In some specific embodiments, the base plate 1 and the intestinal model 2 are an integrated structure.
[0045] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the embodiments. Obviously, the embodiments described are only a part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.
[0046] Example 1
[0047] Combine Figures 1 to 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 to embed intestinal tissue is arranged with the first dehydration hole 3.
[0048] The intestinal model 2 is a spiral structure, and the extension distance of each 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 10mm 2 The height of the intestinal model 2 is 4 mm and the thickness is 0.8 mm.
[0049] 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.
[0050] Example 2
[0051] 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 12mm 2 The height of the intestinal model 2 is 4.5 mm and the thickness is 1 mm.
[0052] Example 3
[0053] 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.
[0054] Example 4
[0055] The difference from Example 1 is that the cross-sectional area of the second dehydration hole 4 in the vertical axis direction is 4mm 2 .
[0056] Comparative Example 1
[0057] Different from Example 1, the first dehydration hole 3 and the second dehydration hole 4 are not provided on the intestinal model 2.
[0058] Comparative Example 2
[0059] The difference from Example 1 is that the cross-sectional area of the first dehydration hole 3 on the intestinal model 2 in the vertical axis direction is 30mm 2 .
[0060] Comparative Example 3
[0061] Different from Example 1, the intestinal model 2 is a spiral structure, and the extension distance of each turn of the spiral structure is 150% of the diameter of the sample to be embedded.
[0062] Test 1
[0063] The molds provided in Example 1 were used to process intestinal tissue samples containing contents. The samples included duodenum tissue, jejunum tissue, ileum tissue, colon tissue, and rectum tissue. After the intestinal tissue was isolated, excess fat tissue on the intestine was trimmed and the tissue of the corresponding intestinal segment was cut. The diameter of the small intestine tissue was about 3 mm, and the diameter of the large intestine tissue was about 3.5 mm. The samples were processed according to the following steps:
[0064] Take the mold and insert the intestinal tissue exactly in the center of Intestinal Model 2. Arrange the intestinal tissue on the model and place it in fixative for 24 hours. Place the fixed tissue and the mold into the finished embedding cassette and dehydrate.
[0065] The program set for the automatic dehydrator is: 10% paraformaldehyde - 2 hours; 75% ethanol - 1 hour; 80% ethanol - 1 hour; 95% ethanol 1--1 hour; 95% ethanol 2--1 hour; anhydrous ethanol 1--30 minutes; anhydrous ethanol 2--30 minutes; xylene 1--30 minutes; xylene 2--30 minutes; paraffin 1--1 hour; paraffin 2--1 hour; paraffin 3--2 hours; the dehydrated tissue is taken out, paraffin embedded, paraffin embedded blocks are prepared, paraffin sections are performed, HE staining is performed, and the section results are obtained.
[0066] like Figure 4Figures a through e show tissues of the duodenum, jejunum, ileum, colon, and rectum, respectively. The intestinal tissues fixed in the mold maintained structural integrity, maintaining a Swiss roll shape. Even after dehydration, they remained intact. The mold, combined with the dehydration process, ensured that all sections of the intestinal tissue, including the contents, remained intact, without compromising histopathological observation.
[0067] Test 2
[0068] The difference from Experiment 1 is that the molds provided in Examples 2 to 4 and Comparative Examples 1 to 3 are used for the following operations: Figure 5 As shown, a through c represent comparative examples 1 through 3, and d through f represent examples 2 through 4. Compared to example 1, the intestinal tissue in examples 2 and 3 exhibited no deformation, and the sections cut out were intact. In example 4, because the cross-sectional area perpendicular to the axis of the second dehydration hole 4 was equal to the cross-sectional area perpendicular to the axis of the first dehydration hole 3, the contact between the solution and the intestinal tissue during the fixation and dehydration processes was affected. While the intestinal tissue exhibited slight deformation, the sections cut out were intact, which did not affect histopathological observation. 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 dehydration reagent is reduced, so the tissue section results show that the tissue is wrinkled and fragmented, affecting the section quality and pathological section observation; in Comparative Example 2, the length of the first dehydration hole 3 is increased, the tissue is deformed during the fixation and dehydration process, and the tissue cannot be embedded on the same cutting surface, affecting the embedding effect; in Comparative Example 3, the diameter of the expansion distance of each circle of the spiral structure is increased, the tissue is deformed during the fixation and dehydration process, and the tissue cannot be embedded on the same cutting surface, affecting the embedding effect.
[0069] Test 3
[0070] The difference from Experiment 1 is that during the fixation process, the jejunum tissue was fixed directly without using a mold, and the slice results were obtained after fixation, dehydration, embedding, and staining. Figure 6 As shown in the figure, during the fixation process, the shape of the intestinal tissue is not fixed. Therefore, after fixation and dehydration, the tissue is not fully embedded during the embedding process due to the limited size of the embedding mold. After staining, the staining results are obtained. Due to the incomplete tissue and the breakage during the embedding process, the tissue is artificially damaged, which increases the difficulty of pathological interpretation.
[0071] 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 above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions 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 vertical axis direction 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).
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 greater than the cross-sectional area of the first dehydration hole (3) in the direction perpendicular to the axis.
4. 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 vertical axis direction is 7 to 12 mm 2 .
5. The intestinal content tissue processing mold according to claim 4, characterized in that: 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) in the vertical axis direction is 10mm 2 .
6. The intestinal content tissue processing mold according to claim 1, characterized in that: The height of the intestinal model (2) is 3.5 to 10 mm.
7. The intestinal content tissue processing mold according to claim 6, characterized in that: The height of the intestinal model (2) is 4 mm.
8. The intestinal content tissue processing mold according to claim 1, characterized in that: The thickness of the intestinal model (2) is 0.5-1 mm.
9. The intestinal content tissue processing mold according to claim 8, characterized in that: The thickness of the intestinal model (2) is 0.8 mm.
10. The intestinal content tissue processing mold according to any one of claims 1 to 9, characterized in that: The base plate (1) and the intestinal model (2) are an integrated structure.