CN114383922A - A kind of standardization operation method of embedding technology - Google Patents

Abstract

The invention provides a standardized operation method for an embedding technology, which belongs to the technical field of biological tissue sample processing, improves the operation standardization of pathological sections of different tissues and the rationality of embedding operation by standardizing pre-embedding processing, embedding operation and post-embedding processing, further improves the quality of the sections, is convenient for effectively diagnosing different pathological tissues, ensures the quality of pathological diagnosis, provides a basis for clinical diagnosis and treatment, carries out comprehensive standardized operation for the conventional histopathological embedding technology, and provides powerful guarantee for improving the embedding quality.

Description

A kind of standardization operation method of embedding technology

technical field

The invention relates to the technical field of biological tissue sample processing, in particular to a standardized operation method of embedding technology.

Background technique

Paraffin section (paraffin section) is the most widely used method in conventional histology preparation technology. It is not only used to observe the morphological structure of normal cell tissue, but also used in cell biology, pathology and medicine to study, observe and judge cells. The main method of group morphological change.

The quality of tissue slicing and whether it is conducive to slicing are affected by many factors and depend on each step of tissue processing, and the quality of each step is closely related to the previous step. While good tissue sectioning requires proper tissue fixation, accurate tissue handling, and delicate tissue sectioning assurance, the step most likely to destroy tissue sections is improper tissue embedding, including improper tissue placement and improper alignment Orientation, the standardization of the embedding operation or not will affect the quality of tissue sections.

Although the "Clinical Technical Operation Standard Pathology Volume" has standardized requirements for tissue block paraffin embedding technology, the specific operation standard details are not perfect. Based on this, the present invention proposes a standardized operation method for embedding technology, which is a routine The histopathological embedding technology carries out comprehensive standardized operation, which provides a strong guarantee for improving the embedding quality.

SUMMARY OF THE INVENTION

In view of the above problems, the present invention provides a standardized operation method of embedding technology.

The object of the present invention adopts the following technical solutions to realize:

A standardized operation method for embedding technology, comprising the following steps: tissue block fixation, dehydration, transparency, dipping in wax, and embedding; wherein, the embedding step specifically includes the following steps:

(1) Preparation before embedding;

Set the embedding machine to the power-on state, turn on the switch of the cold table, check the wax volume of the wax tank, and confirm whether the wax nozzle can discharge the wax normally;

Clean the operating table of the embedding machine to avoid visible tissue debris or foreign bodies on the table;

Prepare enough embedding frames of suitable size and clean tweezers;

Choose an embedding box that matches the tissue block, so that there is a 1mm gap around the tissue block and the bottom edge of the embedding box during embedding;

(2) Tissue block embedding

Inject the wax solution into the embedding frame, the liquid level is flush with the upper edge, and the filling but does not overflow shall prevail, and then transfer the embedding frame filled with the wax solution to the hot stage;

One-time filling can avoid the possibility of tissue dispersal caused by the secondary waxing, reduce the excessive waste of wax liquid caused by the secondary waxing, reduce the working pressure of the hand caused by repeated waxing, and save the waxing time. Improve the embedding speed;

Check the pathological number including the secondary number of the tissue block, the number of blocks and the requirements of the sampling physician for the embedding surface; identify the tissue type according to the technical worksheet;

Use hot clean tweezers to take out the tissue in the embedding box, arrange the tissue according to the plan on the hot table, quickly place the tissue block in the center of the bottom of the wax solution in a fixed position, and lay it flat to ensure that the tissue and wax solution are sufficient. After fusion, move it smoothly to the small cold table of the embedding machine with a small amount of water left. After the bottom of the wax liquid begins to solidify, cover the embedding box and press the embedding mold. After the wax liquid around the embedding box begins to solidify, the The embedding mold is moved to a cold stage to accelerate solidification;

Tissue positioning standard: the embedded tissue is centered;

Tissue aggregation standard: close aggregation, no obvious gap visible to the naked eye;

Tissue embedding level standard: the embedding surface is consistent in height, to ensure that the trimming can be synchronized in place at one time, the thickness of the wax block is consistent, and the tissue blocks are not uneven;

Standard of organization arrangement: the organization blocks are arranged in the same direction and neatly, and the square arrangement is preferred; if the square cannot be arranged, the arrangement should be rectangular and should be in a vertical column;

After embedding one wax block, open the cover of the other embedding box, so that only one wax block to be embedded will always appear on the embedding platform, reducing the time wasted invisibly caused by judging the pathological number when checking the number of tissue blocks; The action of opening the box should be light to avoid the risk of tissue ejection and mutual confusion. It is also convenient for cleaning the embedding platform at any time, reducing the risk of contamination between tissues, and reducing the condensation of tissue blocks caused by temperature drop due to delay in embedding. , thereby affecting the risk of slice quality;

The embedding process should be operated quickly to avoid the solidification of the molten wax before the tissue block is embedded; reduce the unevenness of the high-level tissue caused by the slow embedding speed, which may lead to disintegrated tissue, excessive trimming and damaged tissue, and tissue during slicing. cracks appear between

When embedding, it is necessary to strictly prevent all kinds of foreign body contamination, and do not embed irrelevant tissues such as sutures, paper scraps or other foreign bodies (especially hard foreign bodies) into the wax block;

Adding water to the small cooling table can speed up the cooling speed, avoid the wax liquid sticking to it and cause the cooling to slow down, ensure the cooling surface is even, solve the cooling quality decline caused by the poor cooling function of the small cooling table, and also reduce the wax liquid sticking to the small cooling table. Waste of time to scrape off the wax block on the cold table;

The embedding mold should be pressed as much as possible at one time to reduce the overflow of wax liquid and reduce repetitive actions;

(3) Post-embedding treatment

Take out the solidified embedding wax block from the embedding mold, remove the excess paraffin around the tissue block with a blade, and trim the embedding wax block into a regular square or rectangle based on 1-2mm of paraffin around the tissue block;

Clean the surface of the embedding machine.

Preferably, the largest surface of the tissue block or the tissue surface at the specially designated place is buried in the wax liquid, and the multiple small tissues embedded in the same wax block are close to each other and are located on the same plane;

Embedding surface requires that the maximum cutting surface faces downward for those who have cut surface requirements, and the maximum surface facing downward for those without cutting surface requirements. Clearly, it should be buried vertically, so that the tissue structure of all levels can be seen under the microscope after staining. The kidneys have cortex and medulla.

Preferably, the embedding frame is made of stainless steel or aluminum alloy.

Preferably, the bottom of the inner wall of the embedding chamber of the embedding frame is provided with a concave surface concave downward.

In the slicing process, the embedded wax block needs to be trimmed first. Since the embedded wax block is rectangular, the contact area and times of the blade are increased, which makes the blade easily damaged and easily causes knife marks in the section. In the present invention, a downwardly concave concave surface is arranged at the bottom of the inner wall of the embedding chamber, so that the sliced surface of the wax block has micro-convexities, which reduces the contact area between the wax block and the blade during trimming, thereby protecting the blade.

Preferably, the inner wall of the embedding frame is coated with a Teflon coating.

In order to further reduce the adhesion of the wax block after solidification and make the wax block easier to demould, a Teflon coating can be applied to the inner wall.

Preferably, water that occupies 2/3 of the area of the small cooling table is retained on the small cooling table;

If the water is difficult to retain, you can use wax around the small cold table.

Preferably, the temperature difference between the wax solution used for embedding and the tissue dipping agent is 2°C;

The temperature difference between the embedding paraffin and the tissue dipping agent is 2 degrees, so both of them are in a molten state, so that the tissue and embedding paraffin can be fused together after embedding the condensed wax block. Otherwise, if the temperature of the embedding paraffin is too high Low, paraffin begins to condense during embedding, and it is too late to operate, and it is easy to bring in air bubbles. In this way, after the embedded wax block is condensed, the density of the wax block is often uneven, the tissue and paraffin are separated, and it is difficult to cut a complete section. wax embedded.

Preferably, the tissue block is coarsely stained with eosin in the dehydration process;

Coarse staining of eosin allows easier identification of tissue fragments.

The beneficial effects of the present invention are:

The invention proposes a standardized operation method of embedding technology, which improves the operation standardization of different tissue pathological slices and the rationality of embedding operation, thereby improving slice quality, facilitating effective diagnosis of different pathological tissues, ensuring the quality of pathological diagnosis, and providing clinical benefits for clinical practice. It provides a basis for diagnosis and treatment, and provides a strong guarantee for the comprehensive standardized operation of conventional histopathological embedding technology and the improvement of embedding quality.

Description of drawings

The present invention will be further described by using the accompanying drawings, but the embodiments in the accompanying drawings do not constitute any limitation to the present invention. For those of ordinary skill in the art, under the premise of no creative work, other Attached.

1 is a schematic diagram of an embedding frame provided with a downwardly concave concave surface;

Figure 2 is a standard diagram of the placement of general masses;

Figure 3 is a standard diagram of the placement of fibroids;

Figure 4 is a standard diagram of the placement of the appendix;

Figure 5 is a standard illustration of the placement of the intima;

Figure 6 is a standard illustration of the placement of cysts;

Fig. 7 is the standard diagram of fixed point, location and orientation of the cyst embedding surface;

Fig. 8 is the fixed point, location, orientation standard diagram of the skin embedded surface;

Fig. 9 is the standard diagram of fixed point, location and orientation of the tumor embedding surface;

Fig. 10 is the standard diagram of fixed point, location, and orientation of the endometrium and myometrial embedding surface;

Figure 11 is an illustration of the positioning, positioning, and orientation criteria of the cervical embedding surface.

Detailed ways

The present invention will be further described with reference to the following examples.

The embodiment of the present invention relates to a standardized operation method of embedding technology, which includes the following steps: tissue block fixation, dehydration, transparency, dipping in wax, and embedding; wherein, the embedding step specifically includes the following steps:

(1) Preparation before embedding;

Set the embedding machine to the power-on state, turn on the switch of the cold table, check the wax volume of the wax tank, and confirm whether the wax nozzle can discharge the wax normally;

Clean the operating table of the embedding machine to avoid visible tissue debris or foreign bodies on the table;

Prepare enough embedding frames of suitable size and clean tweezers;

Choose an embedding box that matches the tissue block, so that there is a 1mm gap around the tissue block and the bottom edge of the embedding box during embedding;

(2) Tissue block embedding

Inject the wax solution into the embedding frame, the liquid level is flush with the upper edge, and the filling but does not overflow shall prevail, and then transfer the embedding frame filled with the wax solution to the hot stage;

Check the pathological number including the secondary number of the tissue block, the number of blocks and the requirements of the sampling physician for the embedding surface; identify the tissue type according to the technical worksheet;

Use hot clean tweezers to take out the tissue in the embedding box, arrange the tissue according to the plan on the hot table, quickly place the tissue block in the center of the bottom of the wax solution in a fixed position, and lay it flat to ensure that the tissue and wax solution are sufficient. After fusion, move it smoothly to the small cold table of the embedding machine. The water that occupies 2/3 of the area of the small cold table remains on the small cold table. After the bottom of the wax liquid begins to solidify, cover the embedding box and press the embedding mold. The embedding mold should be pressed as much as possible at one time to reduce the overflow of wax liquid and reduce repeated actions; after the wax liquid around the embedding box begins to solidify, move the embedding mold to a cold table to accelerate solidification;

The largest face of the tissue block or the tissue face at the specially designated place is buried in the wax liquid, and the multiple small tissues embedded in the same wax block are close to each other and are located on the same plane;

Embedding surface requires that the maximum cutting surface faces downward for those who have cut surface requirements, and the maximum surface facing downward for those without cutting surface requirements. Clearly, it should be buried vertically, so that the tissue structure of all levels can be seen under the microscope after staining. The kidneys have cortex and medulla, etc.;

When embedding, it is necessary to strictly prevent all kinds of foreign body contamination, and do not embed irrelevant tissues such as sutures, paper scraps or other foreign bodies (especially hard foreign bodies) into the wax block;

Referring to Figures 1-5, the standard diagrams for the placement of general masses, fibroids, appendix, intima, and cysts are shown in sequence. Referring to Figures 6-10, the positioning, positioning, and orientation standards of the cyst embedding surface are gathered, Side-by-side and centered; the standard of positioning, positioning, and orientation of the skin-embedded surface is that the skin faces to the left; the standard of positioning, positioning, and orientation of the tumor-embedded surface is the center; the standard of positioning, positioning, and orientation of the endometrium and myometrium-embedded surface The standard is longitudinal, side-by-side, and centered; the fixed point, positioning, and orientation of the cervical embedding surface are longitudinal, side-by-side, and centered.

The tissue block is coarsely stained with eosin in the dehydration link;

(3) Post-embedding treatment

Take out the solidified embedding wax block from the embedding mold, remove the excess paraffin around the tissue block with a blade, and trim the embedding wax block into a regular square or rectangle based on 1-2mm of paraffin around the tissue block;

Clean the surface of the embedding machine.

The temperature difference between the embedding paraffin and the tissue dipping agent is 2 degrees, so both of them are in a molten state, so that the tissue and embedding paraffin can be fused together after embedding the condensed wax block. Otherwise, if the temperature of the embedding paraffin is too high Low, paraffin begins to condense during embedding, and it is too late to operate, and it is easy to bring in air bubbles. In this way, after the embedded wax block is condensed, the density of the wax block is often uneven, the tissue and paraffin are separated, and it is difficult to cut a complete section. wax embedded.

Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, but not to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that , the technical solutions of the present invention may be modified or equivalently replaced without departing from the spirit and scope of the technical solutions of the present invention.

Claims (8)

1. An embedding technology standardized operation method is characterized by comprising the following steps: fixing tissue blocks, dehydrating, transparentizing, waxing and embedding; wherein the embedding step specifically comprises the following steps:

(1) preparing before embedding;

the embedding machine is set to be in a starting state, a cold table switch is turned on, the wax quantity of a wax cylinder is checked, and whether a wax nozzle can normally discharge wax is determined;

cleaning the operating table top of the embedding machine to avoid macroscopic tissue debris or foreign bodies from appearing on the table top;

preparing sufficient embedding frames with proper sizes and clean tweezers;

selecting an embedding box matched with the tissue block, and keeping a gap of 1mm between the periphery of the tissue block and the bottom edge of the embedding box during embedding;

(2) tissue block embedding

Injecting the wax liquid into the embedding frame, wherein the liquid level is parallel to the upper edge, and the embedding frame filled with the wax liquid is transferred to a hot table on the basis of filling the embedding frame but not overflowing;

checking the pathological number and the number of the tissue blocks including the secondary number and the requirements of a drawing physician on an embedding surface; identifying the tissue type according to the technical work order;

taking out tissues in the embedding box by using hot clean tweezers, planning the tissue placement on a hot table according to the tissue placement, rapidly placing the tissue blocks into the center of the bottom of the wax liquid in a fixed-point positioning and orienting manner, straightening and paving the tissue blocks to ensure that the tissues and the wax liquid are fully fused, then stably moving the tissue blocks to a small cold table of an embedding machine with a small amount of water, covering the embedding box after the bottom of the wax liquid begins to solidify, pressing an embedding mold, and moving the embedding mold to the cold table to accelerate the solidification after the wax liquid around the embedding box begins to solidify;

(3) post-embedding treatment

Taking out the solidified embedding wax block from the embedding mould, removing excessive paraffin wax around the tissue block by using a blade, and trimming the embedding wax block into a regular square or rectangle by taking paraffin wax of 1-2mm reserved around the tissue block as a standard;

cleaning the operating table top of the embedding machine.

2. An embedding technique standardization operating method as claimed in claim 1, wherein the largest face of the tissue block or the specially designated tissue face is embedded downwards into the wax liquid, and a plurality of small tissues embedded in the same wax block are close to each other and located on the same plane.

3. The standardized embedding technique operating method as claimed in claim 1, wherein the embedding frame is made of stainless steel or aluminum alloy.

4. The standardized embedding technique operating method as claimed in claim 1, wherein the bottom of the inner wall of the embedding chamber of the embedding frame is provided with a concave surface which is concave downwards.

5. An embedding technique standardized operation method as claimed in claim 1, characterized in that the inner wall of the embedding frame is coated with teflon coating.

6. The standardized embedding technique operating method as claimed in claim 1, wherein the small cooling stage is provided with water occupying 2/3 of the area of the small cooling stage.

7. An embedding technique standardization operating method as claimed in claim 1, wherein the temperature difference between the embedding wax liquid and the tissue waxing agent is 2 ℃.

8. The standardized manipulation method of embedding technique as claimed in claim 1, wherein the tissue block is stained with eosin in a dehydration step.

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