Puncture tissue embedding method and application

Abstract

The invention discloses a puncture tissue embedding method and application. The embedding method comprises the following steps: covering the surface of a puncture tissue with OCT embedding glue, and turning over the puncture tissue until the puncture tissue is completely wrapped by the OCT embedding glue; then the puncture tissue wrapped with the OCT embedding adhesive is clamped out and placed on a platform without the OCT embedding adhesive; then the puncture tissue is flatly laid on gasket paper and pressed, it is guaranteed that the puncture tissue and the gasket paper are completely attached, and the surface of the puncture tissue is covered with OCT embedding glue; the gasket paper padded with the puncture tissue is placed in a clean embedding groove, the periphery of the gasket paper is slightly pressed, and the gasket paper is attached to the bottom of the embedding groove; oCT embedding glue is extruded to fill the embedding groove, the embedding groove is put into a pre-cooled freezing embedding instrument, a freezing support is covered, a heat absorption hammer is pressed, and timing is conducted; and after timing is finished, taking out the freezing support to finish embedding. The method is simple to operate, the punctured tissue can be obviously seen after embedding, the tissue can be completely cut conveniently, and the frozen section is facilitated.

Description

Technical Field

[0001] The invention belongs to the field of inspection and detection, and particularly relates to an embedding method and application of punctured tissue. Background Art

[0002] Biopsy is an important method for diagnosing diseases. The open incision method can obtain more living tissue, which is conducive to the development of detection, but the patient's recovery period is long and the risk of infection is high. The puncture biopsy method has a small incision and obtains less living tissue, which makes the detection more difficult, but it is conducive to the patient's recovery and has a higher safety factor.

[0003] Intraoperative frozen pathology diagnosis is an emergency consultation with a pathologist during the operation when the surgeon is performing an operation and asks the pathologist to diagnose the disease related to the surgical plan. The issuance of an intraoperative frozen pathology diagnosis report requires sampling, frozen embedding, frozen sectioning, frozen HE staining and observation under a microscope; among them, the thickness of conventional tissue freezing sampling is 3-4mm, and when frozen embedding, it only needs to be embedded according to the required cutting surface. If it is slightly uneven, the tissue can be completely cut or the lesion can be completely cut through deep cutting; but the diameter of the punctured tissue is small, and uneven embedding will lead to incomplete cutting, which may lead to missed diagnosis. Therefore, conventional frozen embedding methods are not suitable for punctured tissue.

[0004] At present, there are only two embedding methods for frozen embedding of punctured tissues, which are respectively called puncture tissue embedding operation 1 and puncture tissue embedding operation 2 for easy distinction. Puncture tissue embedding operation 1 is to freeze the sample head with cryoembedding gel, wait for the cryoembedding gel to completely solidify, put it into the cryostat to trim a flat surface, the function of the flat surface is to provide a flat support for the puncture strip, mark the relative position of the sample head, take out the sample head, and then lay the puncture tissue strips on the flat surface, then add cryoembedding gel to the puncture strip, wait for it to solidify, put the sample head back to the marked position, and then perform rough trimming, fine trimming and slice production. The second operation of puncture tissue embedding is to freeze the sample head with cryoembedding gel, wait until the cryoembedding gel is in a translucent semi-solidified state, use a freezing hammer to press on the cryoembedding gel, and the cryoembedding gel is compressed to freeze a plane, and then lay the punctured tissue strips on the plane, and then evenly apply a layer of cryoembedding gel on the puncture strips until the specimen is completely covered, and gently press it flat with a freezing hammer, and then slice it. Both of these embedding methods have obvious defects: the first defect is that during the embedding process, when the punctured tissue is placed on the low-temperature plane formed by the frozen section embedding agent on the sample head, the contact part will solidify instantly and cannot be moved when the fresh punctured tissue contacts the low-temperature plane, which may cause the punctured tissue to be embedded unevenly, such as an "arch bridge" shape, with both ends fixed and the middle arched. When slicing, the arched part will be trimmed off and the slice cannot be made; the second defect is that the plane provided by the solidified embedding glue on the sample head is used to support the punctured tissue, and the diameter of the punctured tissue is not uniform, which causes the bottom surface of the punctured tissue to be flat but the cross-section to be uneven. If the punctured tissue is to be cut completely, a lot of tissue will be lost; the third defect is that after these two embedding methods flatten the punctured tissue, frozen embedding glue must be added to the punctured tissue as soon as possible, otherwise the punctured tissue will not be able to fuse with the subsequently added embedding glue after solidification, and the punctured tissue will be separated from the subsequently added embedding glue and cannot be sliced.

[0005] Therefore, it is necessary to develop an embedding technology that is more conducive to puncture tissue preparation. Summary of the invention

[0006] The primary purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and to provide a method for embedding punctured tissue.

[0007] Another object of the present invention is to apply the embedding method of the punctured tissue to frozen sections.

[0008] The purpose of the present invention is achieved through the following technical solution: A method for embedding punctured tissue comprises the following steps:

[0009] (1) Precool the freezing embedding apparatus to below -20°C and clean the embedding tank;

[0010] (2) Place the punctured tissue on the platform, cover the surface of the punctured tissue with OCT embedding gel, and turn the punctured tissue over until it is completely wrapped by the OCT embedding gel; then remove the punctured tissue wrapped with OCT embedding gel and place it on a platform without OCT embedding gel to reduce the amount of OCT embedding gel wrapped in the punctured tissue;

[0011] (3) Lay the punctured tissue obtained in step (2) flat on the gasket paper, press to ensure that the punctured tissue and the gasket paper are completely in contact, and cover the surface of the punctured tissue with OCT embedding glue;

[0012] (4) placing the spacer paper with the punctured tissue obtained in step (3) into the embedding tank, lightly pressing the spacer paper around to make the spacer paper fit the bottom of the embedding tank; squeezing out the OCT embedding glue to fill the embedding tank, and placing it in a pre-cooled freezing embedding instrument; covering the freezing tray, pressing the endothermic hammer, and timing;

[0013] (5) When the timer ends, remove the frozen tray and embedding is complete.

[0014] The precooling temperature in step (1) is preferably -30°C to -40°C; more preferably -35°C.

[0015] The punctured tissue described in step (2) includes the punctured tissue obtained by coarse needle puncture and the punctured tissue obtained by fine needle puncture.

[0016] The platform described in step (2) is a positioning shovel, a photographing board, or other clean flat surface.

[0017] The spacer paper described in step (3) is a light-transparent and soft thin paper, which is convenient for observation and does not damage the blade during slicing; preferably, the spacer paper has a diameter of 3 cm.

[0018] The pressing in step (3) is preferably performed using tweezers.

[0019] The amount of OCT embedding glue used in step (3) is preferably just enough to cover the punctured tissue.

[0020] The timing duration in step (4) is preferably 25 to 35 seconds, and more preferably 30 seconds.

[0021] The application of the above-mentioned embedding method of punctured tissue in the preparation of frozen tissue sections.

[0022] Compared with the prior art, the present invention has the following advantages and effects:

[0023] (1) The present invention uses a freezing embedding apparatus, which can reduce the formation of tissue ice crystals and improve the quality of slices.

[0024] (2) The present invention wraps the punctured tissue with cryoembedding gel in advance, thereby avoiding the punctured tissue and the cryoembedding gel from separating and thus being unable to prepare slices.

[0025] (3) The present invention uses small sample paper as a support, and the position of the punctured tissue can be placed at will.

[0026] (4) The present invention uses small sample gasket paper for leveling, which can make the punctured tissue section flat and avoid missed diagnosis due to incomplete cutting.

[0027] (5) The embedded tissue of the present invention has a smooth cross section, which reduces the waste of punctured tissue and retains more tissue for the final diagnosis after freezing in paraffin. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a photograph of the punctured tissue obtained by coarse needle puncture covered with OCT embedding gel.

[0029] Figure 2 This is a photograph of the removal of excess OCT embedding gel around the punctured tissue obtained by thick needle puncture.

[0030] Figure 3 This is a photograph of the puncture tissue obtained by coarse needle puncture placed on a small sample spacer paper.

[0031] Figure 4 Here is a photo of a small sample of shim paper.

[0032] Figure 5 This is a photograph of placing a small sample pad paper containing the puncture tissue obtained by coarse needle puncture into the embedding tank of the freezing embedding instrument.

[0033] Figure 6 This is a photo of the embedding tank being covered with a freezing tray and pressed with a heat absorbing hammer.

[0034] Figure 7 This is a photograph of the punctured tissue obtained by thick needle puncture after embedding.

[0035] Figure 8 This is a cross-sectional view of the punctured tissue obtained by coarse needle puncture.

[0036] Fig. 9 This is a photograph of the slide obtained after staining of the punctured tissue section obtained by coarse needle puncture.

[0037] Fig.10 This is a photograph of the punctured tissue obtained by fine needle puncture.

[0038] Fig.11 This is a photograph of the punctured tissue obtained by fine needle puncture covered with OCT embedding gel.

[0039] Fig.12This is a photograph of the removal of excess OCT embedding gel around the punctured tissue obtained by fine needle puncture.

[0040] Fig.13 This is a photograph of a small sample pad of tissue obtained by fine needle aspiration.

[0041] Fig.14 This is a photograph of OCT embedding gel filling the embedding groove containing the punctured tissue and small sample spacer paper.

[0042] Fig.15 This is a photograph of the punctured tissue obtained by fine needle puncture after embedding.

[0043] Fig.16 This is a cross-sectional view of the punctured tissue obtained by fine needle puncture.

[0044] Fig.17 This is a photograph of the slide obtained after staining of the puncture tissue section obtained by fine needle puncture. DETAILED DESCRIPTION

[0045] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0046] Example 1

[0047] A method for embedding puncture tissue obtained by thick needle puncture, comprising the following steps:

[0048] (1) Precool the cryo-embedding apparatus (Milestone, Italy) to -35°C and clean the embedding tank;

[0049] (2) Place the puncture strip on the positioning spatula, squeeze a small amount of OCT embedding gel on the surface of the punctured tissue, and use tweezers to flip the puncture strip until it is completely covered with OCT embedding gel (such as Figure 1 Then, each puncture strip is removed by clamping to reduce the amount of OCT embedding glue wrapped in the puncture strip (as shown in Figure 2 As shown), the purpose of this step is to wrap the punctured tissue with embedding glue to facilitate smooth fusion with the subsequent embedding glue; less embedded glue is easy to fit the punctured tissue to the small sample gasket paper as much as possible;

[0050] (3) Take out the puncture strip and lay it flat on the small sample gasket paper. Use tweezers to press appropriately to ensure that the punctured tissue and the small sample gasket paper are completely in contact (such as Figure 3 As shown in the figure), squeeze out a little OCT embedding gel to cover the surface of the puncture strip; small sample spacer paper as shown in the figure Figure 4 As shown, it is a light blue thin sheet of paper with a diameter of 30 mm;

[0051] (4) Place the small sample spacer paper with the punctured tissue obtained in step (3) into the embedding tank of the cryo-embedding instrument, gently press the small sample spacer paper around to make the small sample spacer paper fit the bottom of the embedding tank, and then quickly squeeze out the OCT embedding glue to fill the embedding tank (such as Figure 5 Cover the freezing tray and press the heat absorbing hammer (as shown); Figure 6 As shown), time 30s;

[0052] (5) When the timer is up, take out the cryo-tray and embedding is complete. The embedded punctured tissue is shown in Figure 7. The shape of the punctured tissue can be seen through the small sample spacer paper. Put it into the cryo-tome, adjust the direction of the chuck, and trim it carefully to cut the whole tissue. Figure 8 The frozen sections obtained are as follows Fig. 9 shown.

[0053] Example 2

[0054] A method for embedding puncture tissue obtained by fine needle puncture, comprising the following steps:

[0055] (1) Precool the freezing embedding apparatus to -35°C and clean the embedding tank;

[0056] (2) Place the puncture strip on the camera plate ( Fig.10 ), squeeze a small amount of OCT embedding glue on the surface of the punctured tissue ( Fig.11 ), use tweezers to turn the puncture strips until they are completely wrapped by OCT embedding gel, and then take out each puncture strip to reduce the amount of OCT embedding gel wrapped in the puncture strips ( Fig.12 );

[0057] (3) Clip out the puncture strip and lay it flat on the small sample pad paper ( Fig.13 ), squeeze out a little OCT embedding gel to cover the surface of the puncture strip;

[0058] (4) Clamp the small sample gasket paper and place it in the embedding tank of the freezing embedding instrument. First, gently press the punctured tissue to ensure that the small sample gasket paper fits the punctured tissue. Then gently press the small sample gasket paper around to make the small sample gasket paper fit the bottom of the embedding tank. Then quickly squeeze out the OCT embedding glue to fill the embedding tank ( Fig.14 ), cover with freezing tray, press on endothermic hammer, and time for 30s;

[0059] (5) When the timer is up, take out the frozen tray and embedding is complete. Fig.15 As shown, the shape of the punctured tissue can be seen through the small sample spacer paper; put it into the freezing slicer, adjust the direction of the chuck, and finely trim to cut the entire tissue, such as Fig.16 The frozen sections obtained are as follows Fig.17 shown.

[0060] It can be seen that compared with the two existing embedding methods for punctured tissue, the present invention has the following advantages: the first advantage is that the punctured tissue is pre-wrapped with a frozen section embedding agent instead of directly placing the punctured tissue (such as punctured tissue embedding operation 1 and punctured tissue embedding operation 2), thereby avoiding the separation of the punctured tissue and the subsequently added frozen section embedding agent, and the slicing can be carried out smoothly; the second advantage is that before being placed in the embedding slot of the frozen embedding instrument, the punctured tissue can be placed at an arbitrary position to prevent the punctured tissue from crossing after embedding, which may cause the punctured tissue to be unable to be completely cut, while the existing punctured tissue embedding method, due to the use of a frozen section embedding agent in advance, The sample head is wrapped, so when the punctured tissue is placed on the low-temperature plane formed by the frozen section embedding agent on the sample head, the contact part will solidify instantly when the fresh punctured tissue contacts the low-temperature plane, making it impossible for the punctured tissue to move and adjust its position. After placement, the frozen section embedding agent must be added quickly, and the embedding effect depends on the proficiency of the operator; the third advantage is that the punctured tissue can be clearly seen after embedding, which is convenient for cutting the entire tissue; the fourth advantage, which is also the biggest advantage, is that the punctured tissue can be embedded flat, which is conducive to cutting the entire punctured tissue and reducing tissue waste, leaving more tissue for the final diagnosis after freezing in paraffin.

[0061] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A method for embedding punctured tissue, characterized in that The following steps are involved: (1) Precool the freezing embedding apparatus to below -20°C and clean the embedding tank; (2) Place the punctured tissue on the platform, cover the surface of the punctured tissue with OCT embedding gel, and turn the punctured tissue over until it is completely wrapped by the OCT embedding gel; then remove the punctured tissue wrapped with OCT embedding gel and place it on a platform without OCT embedding gel to reduce the amount of OCT embedding gel wrapped in the punctured tissue; (3) Lay the punctured tissue obtained in step (2) flat on the gasket paper, press to ensure that the punctured tissue and the gasket paper are completely in contact, and cover the surface of the punctured tissue with OCT embedding glue; (4) placing the spacer paper with the punctured tissue obtained in step (3) into the embedding tank, and gently pressing the spacer paper around to make the spacer paper fit the bottom of the embedding tank; Squeeze out OCT embedding glue to fill the embedding tank, and place it in the pre-cooled freezing embedding instrument; cover the freezing tray, press the endothermic hammer, and start timing; (5) When the timer ends, remove the frozen tray and embedding is complete.

2. The embedding method of punctured tissue according to claim 1, characterized in that: The precooling temperature in step (1) is -30°C to -40°C; The punctured tissue described in step (2) includes the punctured tissue obtained by coarse needle puncture and the punctured tissue obtained by fine needle puncture.

3. The embedding method of punctured tissue according to claim 2, characterized in that: The precooling temperature in step (1) is -35°C.

4. The embedding method of punctured tissue according to claim 1, characterized in that: The platform described in step (2) is a positioning shovel, a photographing board or other clean platform.

5. The embedding method of punctured tissue according to claim 1, characterized in that: The gasket paper in step (3) is a light-transmitting and soft thin paper; The pressing described in step (3) is performed using tweezers.

6. The embedding method of punctured tissue according to claim 1, characterized in that: The amount of OCT embedding glue used in step (3) is just enough to cover the punctured tissue.

7. The embedding method of punctured tissue according to claim 1, characterized in that: The timing described in step (4) is 25 to 35 seconds.

8. The embedding method of punctured tissue according to claim 7, characterized in that: The timing described in step (4) is 30 seconds.

9. Use of the embedding method for punctured tissue according to any one of claims 1 to 8 in preparing frozen tissue sections.

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