Mouse intestinal tract longitudinal cutting table

Abstract

The utility model discloses a mouse intestinal tract longitudinal cutting table which comprises a base and a cover, and the cover covers the base. The two ends of the base are each provided with a protruding stick frame, and a plurality of grooves are formed in each stick frame at equal intervals and correspond to the grooves in the table top of the base in a one-to-one mode. The cover is provided with a plurality of through grooves, and the part, between every two adjacent through grooves, of the cover forms a cutting guide edge. The mouse intestinal tract longitudinal cutting table is used for longitudinally cutting the intestinal tract tissue of a mouse, the operation is simple and rapid, the intestinal tract tissue longitudinal cutting effect is good, and the time from in-vitro to fixation of the intestinal tissue can be effectively shortened.

Description

Technical Field

[0001] The utility model relates to the technical field of mouse intestinal tissue processing, in particular to a mouse intestinal longitudinal cutting table. Background Art

[0002] How to properly handle mouse intestinal tissue, including rapid dissection and timely fixation, is crucial for the subsequent staining of tissue sections. Currently, the commonly used methods for preparing mouse intestinal tissue sections include intestinal rolls and intestinal segments. The main steps of intestinal roll making are: cutting the mouse intestine along the long axis, then rolling and winding the intestine along the cut direction into a roll, and then embedding it into a tissue block. Because of its morphological characteristics similar to Swiss rolls, this technology is also called "Swiss Roll". Intestinal segments reflect some random areas of the entire length of the intestine, while Swiss rolls can reflect all the characteristics of the entire length of the intestine on a single slice, allowing us to conduct detailed and extensive examinations of the intestine. This feature is of great significance in oncology research. Since Magnus proposed the concept of Swiss rolls in 1937 (he used clinical gastric tissue specimens), Reilly and Kirsner first applied it to the intestine in 1965, but the details were insufficient. In 1981, Moolenbeek made the first detailed summary of the preparation of mouse intestinal rolls, and in 1987, Park further expanded this technology to the gastrointestinal tract, esophagus, reproductive system of rats, and large blood vessels and trachea of ​​New Zealand white rabbits. In the past decade, there have been articles intermittently publishing improved Swiss roll technology to prepare better mouse intestinal tissue for immunohistochemistry or immunohistofluorescence staining.

[0003] A key step in preparing mouse intestinal rolls is to cut the intestinal tissue longitudinally. In all the literature related to this step, the experimenters choose to use dissecting scissors to cut the entire length of the mouse intestine. This operation requires the experimenters to have certain anatomical experience and skills, otherwise the edges of the longitudinally cut intestinal tissue are often uneven or even missing, which increases the difficulty of subsequent operations;

[0004] Currently, paraffin embedding is mostly used to prepare mouse intestinal rolls. After we divide the intestinal tissue into segments (for example, the small intestine is divided into three segments, and the large intestine is one segment), each segment of the intestinal tissue is prepared into an intestinal roll. Therefore, four intestinal rolls will be prepared from the full length of the intestine of a mouse, which undoubtedly increases the space load for sample preservation. Summary of the invention

[0005] In view of the above technical deficiencies, the purpose of the utility model is to provide a mouse intestinal longitudinal cutting table; use the longitudinal cutting table instead of dissecting scissors to longitudinally cut the intestinal tissue; after the intestinal tissue of a mouse is segmented (the small intestine is divided into three sections, and the large intestine is one section), prepare it into an intestinal roll frozen tissue block.

[0006] In order to solve the above technical problems, the utility model adopts the following technical solutions:

[0007] The utility model provides a mouse intestinal longitudinal cutting table, which is characterized in that it comprises a base and a cover, wherein the cover covers the base; a raised stick frame is respectively arranged at both ends of the base, a plurality of grooves are equidistantly arranged on the stick frame, and correspond one to one with the grooves on the base table surface; a plurality of through grooves are arranged on the cover, and the cover part between two adjacent through grooves forms a knife cutting guide edge; the upper part of the cross section of the knife cutting guide edge is a trapezoidal structure, and the lower part is a rectangular structure.

[0008] The utility model also relates to a method for making a mouse full-length intestinal frozen Swiss roll by using the mouse intestinal longitudinal cutting table, which is characterized by comprising the following steps:

[0009] Step 1: Sampling and fixation of mouse intestines;

[0010] (1) Sacrifice the mouse by cervical dislocation, shave the mouse's abdominal hair, make a small "V"-shaped incision at the midline of the abdomen, and continue to cut the skin upward to the sternum and downward to the pubic bone to extend the surgical incision;

[0011] (2) After fully exposing the abdominal cavity, find the junction between the stomach and esophagus and cut it slightly above;

[0012] (3) Using the cut end as the starting point, free the gastrointestinal tract from the abdominal cavity and peel off the related mesentery and connective tissue. During the freeing process, pay attention to timely flushing the gastrointestinal tract freed from the abdominal cavity with cold PBS to keep it moist.

[0013] (4) After a complete gastrointestinal tract is freed from the abdominal cavity, it is immediately immersed in cold PBS;

[0014] (5) Use dissecting scissors to divide the small intestine into three equal sections according to length. From the stomach side to the cecum side, mark them as SB1, SB2, and SB3. After cutting off the cecum, the remaining large intestine is marked as LB.

[0015] (6) Use a simple flushing tube filled with cold PBS to flush the intestinal contents; the body of the simple flushing tube used is a 10 ml syringe, and the tip of the tube is made by cutting off the needle of a scalp needle;

[0016] (7) After the intestinal contents are flushed, use the mouse intestinal longitudinal sectioning table to perform longitudinal sectioning of the intestine; first, soak the cut absorbent paper a with cold PBS and place it on the table top of the base; then, pass four stainless steel rods through the intestinal cavity of SB1, SB2, SB3 and LB in turn, and then fix them on the table top and place the lid;

[0017] (8) Hold the front end of the intestine with forceps in one hand and hold a sterile scalpel in the other hand. Use the side wall in the middle of the lid as a fulcrum to cut the intestine evenly along the longitudinal axis. If the intestine is still partially attached to the stainless steel rod, use a 10 ml syringe filled with cold PBS to spray liquid along the direction of the scalpel cut so that the intestine is flat on absorbent paper a.

[0018] (10) After removing the stainless steel rod, moisten another piece of absorbent paper b with cold PBS and cover the cut intestinal tissue with it; roll up the two completely matched absorbent papers along their long axis to form a hollow tube, place it in a 50 ml centrifuge tube filled with 4% PFA, and fix it at room temperature for 24 hours;

[0019] (11) After fixation for 24 hours, PFA was replaced with 15% sucrose solution and dehydrated for 24 hours;

[0020] (12) After dehydration with 15% sucrose for 24 hours, replace it with 30% sucrose solution and continue dehydration for another 24 hours;

[0021] Step 2: Prepare frozen tissue blocks of full-length mouse intestinal rolls;

[0022] (1) After dehydration with 30% sucrose for 24 hours, take out the absorbent paper from the 50 ml centrifuge tube and spread it flat on the table with absorbent paper b facing upwards;

[0023] (2) Carefully lift up the absorbent paper b, hold the front end of the intestine with curved forceps in one hand, and gently hold the farthest end of the intestine with another pair of forceps or bare hands in the other hand. With the intestinal mucosa facing inward, roll up the intestine into a tube shape. Then, use a 30G needle to penetrate the entire intestinal roll slightly inward from the outermost end of the intestine to prevent it from unraveling. Carefully loosen the curved forceps to remove it from the fixed intestinal roll.

[0024] (3) Carefully remove the 30G needle and place the four intestinal rolls upright in order in the tissue embedding box;

[0025] (4) Wrap the four intestinal rolls completely with OCT, and then place them on the quick-freezing table of the freezing slicer for quick freezing and embedding; after the OCT turns white and hardens, they can be sliced ​​or stored in a -80℃ refrigerator for later use.

[0026] The beneficial effects of the utility model are: 1. The mouse intestinal tissue is longitudinally cut by using the "mouse intestinal longitudinal cutting table", which is simple and fast to operate, has a good intestinal tissue longitudinal cutting effect, and can effectively shorten the "time from intestinal tissue ex vivo to fixation" (the mouse intestine will undergo rapid changes in the short time window after sampling and before fixation, and the longer the interval time, the more unfavorable it is for subsequent tissue staining analysis);

[0027] 2. Preparing the intestinal tissue of a mouse into a frozen tissue block can effectively save the storage space of the sample; although paraffin sections can be stored at room temperature, the staining steps are cumbersome (frozen sections do not require pre-steps such as dewaxing), and the effect of immunofluorescence staining is inferior to frozen sections; staining on one section to observe the overview of the full-length intestinal tissue of a mouse allows researchers to conduct a comprehensive and careful observation of the entire intestine, thereby making a systematic and scientific histological analysis. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] In order to more clearly illustrate the embodiments of the utility model or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the utility model. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0029] Figure 1 A schematic diagram of the three-dimensional structure of the base of the mouse intestinal longitudinal cutting table provided by the utility model;

[0030] Figure 2 A schematic diagram of the three-dimensional structure of the mouse intestinal longitudinal cutting table cover provided by the utility model;

[0031] Figure 3 A front view of the lid of the mouse intestinal longitudinal cutting table provided by the utility model;

[0032] Figure 4 for Figure 3 Schematic diagram of the cross-sectional structure in the HH direction;

[0033] Explanation of the reference numerals: 1. base; 2. stick holder; 3. groove; 4. knife cutting guide edge; 5. cover; 6. through slot. DETAILED DESCRIPTION

[0034] The following will be combined with the drawings in the embodiments of the utility model to clearly and completely describe the technical solutions in the embodiments of the utility model. Obviously, the described embodiments are only part of the embodiments of the utility model, not all of the embodiments. Based on the embodiments in the utility model, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the utility model. Example 1

[0035] The method for preparing a mouse whole-length intestinal frozen Swiss roll comprises the following steps:

[0036] Step 1: Sampling and fixation of mouse intestines;

[0037] (1) Sacrifice the mouse by cervical dislocation, shave the mouse's abdominal hair, make a small "V"-shaped incision at the midline of the abdomen, and continue to cut the skin upward to the sternum and downward to the pubic bone to extend the surgical incision;

[0038] (2) After fully exposing the abdominal cavity, find the junction between the stomach and esophagus and cut it slightly above;

[0039] (3) Using the cut end as the starting point, free the gastrointestinal tract from the abdominal cavity, and peel off the related mesentery and connective tissue. During the freeing process, pay attention to timely rinse the gastrointestinal tract freed from the abdominal cavity with cold PBS to keep it moist;

[0040] (4) After a complete gastrointestinal tract is freed from the abdominal cavity, it is immediately immersed in cold PBS;

[0041] (5) Use dissecting scissors to divide the small intestine into three equal sections by length, from the stomach side to the cecum side, and label them SB1, SB2, and SB3. After cutting off the cecum, the remaining large intestine is labeled LB;

[0042] (6) Use a simple flushing tube filled with cold PBS to flush the intestinal contents. The body of the simple flushing tube used is a 10 ml syringe, and the tip of the tube is made by cutting off the needle of a scalp needle;

[0043] (7) After the intestinal contents are flushed, use the "mouse intestinal longitudinal sectioning table" to perform longitudinal sectioning of the intestine. First, soak the cut absorbent paper a with cold PBS and place it on the table of base 1. Then, four stainless steel rods are passed through the intestinal cavity of SB1, SB2, SB3 and LB in turn, and then fixed on the table and lid 5 is placed;

[0044] (8) Hold the front end of the intestine with forceps in one hand and hold a sterile scalpel in the other hand. Use the side wall in the middle of the cover 5 as a fulcrum to cut the intestine open along the longitudinal axis.

[0045] (9) If the intestine is still partially attached to the stainless steel rod, use a 10 ml syringe filled with cold PBS to spray liquid along the direction of the scalpel cut to make the intestine flat on absorbent paper a;

[0046] (10) After removing the stainless steel rod, wet another piece of absorbent paper b with cold PBS and cover it on the cut intestinal tissue. Roll up the two completely matched absorbent papers (the longitudinally cut intestinal tissue is sandwiched between them) along their long axis to form a hollow tube, place it in a 50ml centrifuge tube filled with 4% PFA, and fix it at room temperature for 24 hours;

[0047] (11) After fixation for 24 hours, PFA was replaced with 15% sucrose solution and dehydrated for 24 hours;

[0048] (12) After dehydration for 24 hours with 15% sucrose solution, replace it with 30% sucrose solution and continue dehydration for another 24 hours.

[0049] Step 2: Prepare frozen tissue blocks of full-length mouse intestinal rolls;

[0050] (1) After dehydration with 30% sucrose for 24 hours, take out the absorbent paper from the 50 ml centrifuge tube and spread it flat on the table with absorbent paper b facing upwards;

[0051] (2) Carefully lift up the absorbent paper b, hold the front end of the intestine with a pair of curved forceps in one hand, and gently hold the farthest end of the intestine with another pair of forceps or bare hands in the other hand. With the intestinal mucosa facing inward, roll up the intestine into a tube (the roll). Then use a 30G needle to penetrate the entire intestinal roll slightly inward from the outermost end of the intestine to prevent it from unraveling. Carefully loosen the curved forceps and pull it out of the fixed intestinal roll;

[0052] (3) Carefully remove the 30G needle and place the four intestinal rolls upright in order in the tissue embedding box (SB1 in the upper left corner, SB2 in the upper right corner, SB3 in the lower left corner, and LB in the lower right corner);

[0053] (4) Wrap the four intestinal rolls completely with OCT (avoid bubbles), and then place them on the quick-freezing table of the freezing microtome for quick freezing and embedding. After the OCT turns white and hardens, it can be sliced ​​or stored in a -80℃ refrigerator for later use.

[0054] like Figure 1-4 As shown, the mouse intestine longitudinal cutting table used in the above-mentioned method for making the mouse full-length intestine frozen Swiss roll includes a base 1 and a cover 5, and the cover 5 is covered on the base 1; a raised stick frame 2 is provided at each end of the base 1, and a plurality of grooves 3 are equidistantly provided on the stick frame 2, and correspond one to one with the grooves 3 on the surface of the base 1; a plurality of through grooves 6 are provided on the cover 5, and the part of the cover 5 between two adjacent through grooves 6 forms a knife cutting guide edge 4; the upper part of the cross-section of the knife cutting guide edge 4 is a trapezoidal structure, and the lower part is a rectangular structure.

[0055] Steps:

[0056] First, soak the cut absorbent paper a with cold PBS, and spread it flat on the table top of the base 1, so that the absorbent paper just covers the table top between the stick racks 2 at both ends.

[0057] Insert the stainless steel rod into the cleaned mouse intestine, then insert the two ends of the stainless steel rod into the grooves 3 of the rod racks 2 on both sides, and cover them with a lid 5.

[0058] After that, hold one end of the intestinal tube with forceps in one hand, hold a sterile scalpel in the other hand, make the blade close to the side wall of the cutting guide edge 4 in the middle of the cover 5, and the tip of the blade against the stainless steel stick, and cut the intestine along the longitudinal axis along the stainless steel stick. After cutting, remove the cover 5. If the intestine is still partially attached to the stainless steel stick, use a 10ml syringe filled with cold PBS to spray liquid along the cutting direction of the scalpel to free the intestinal cavity surface and the stainless steel stick and unfold it.

[0059] After removing the stainless steel rod, the intestine is now flat on absorbent paper a, with the intestinal cavity facing up and the serosal surface facing down and attached to absorbent paper a. Wet another piece of absorbent paper b with cold PBS and cover it on the intestinal cavity. Finally, roll up the two completely matched absorbent papers along the long axis of the intestine to form a hollow tube.

[0060] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A mouse intestinal longitudinal cutting table, which has the following characteristics: when in use, the lid is covered on the base; a raised stick rack is provided at each end of the base, and a plurality of grooves are equidistantly provided on the stick rack, and correspond one to one with the grooves on the base surface; a plurality of through grooves are provided on the lid, and the lid portion between two adjacent through grooves forms a knife cutting guide edge.

2. The mouse intestinal longitudinal sectioning machine according to claim 1, characterized in that: The upper part of the cross section of the knife-cutting guide edge is a trapezoidal structure, and the lower part is a rectangular structure.

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