Methods for finding target substances that can be shed on the surface of tissue blocks

Abstract

The present invention discloses a method for detecting a target substance that can be detached from the surface of a tissue block, comprising the following steps: 1) tightly imprinting a tissue block containing the target substance onto the surface of a deformable material and then removing the block; 2) closely contacting the deformable material with the target substance retained on its surface with an absorptive glass slide; and 3) treating the glass slide with the target substance adsorbed thereon to allow the target substance to be detected or observed. This method is simple to operate and can be used for rapid intraoperative pathological diagnosis. It is also highly suitable for tissue samples with uneven surfaces or dispersed lesions, and has great clinical application prospects.

Description

Technical Field

[0001] The invention relates to a sampling method, in particular to a method for searching for detachable target substances on the surface of a tissue block. Background Art

[0002] Intraoperative pathological diagnosis is a method used during surgery to quickly understand the nature of a lesion, providing a diagnostic basis for further surgical resection. Typically, the surgeon removes a small piece of tissue from the patient's lesion and sends it to the pathology department for frozen section preparation for intraoperative pathological diagnosis. However, frozen section preparation is time-consuming, and to understand the pathology from different directions, it is necessary to section all sides of the tissue. Some lesions are large and dispersed, making it difficult for the doctor to obtain a small section from all suspicious areas. For example, skin cancer specimens often consist of a large mass of tissue containing thick subcutaneous fat, which may contain multiple tumor lesions. However, this fat tissue is often bumpy, making it difficult for the pathologist to determine the tissue margins and further dissect the fat, making frozen section preparation difficult. Even if a small piece of tissue can be removed, the time constraints of intraoperative pathology make it difficult to sample and prepare frozen sections from all suspicious areas for analysis.

[0003] In fact, some diagnostic targets do not require a large amount to make a qualitative diagnosis. For example, tumor cells can be judged based on their morphology under a microscope, and some cells with specific properties can be judged through immunostaining. At the same time, targets such as cells can be detached from tissue blocks in small quantities, which provides a novel idea for rapid diagnosis during surgery.

[0004] In order to quickly and comprehensively understand the condition of the entire sample, it is best to sample all surfaces of the entire sample at once. For those tissues with uneven surfaces, it is also hoped to collect detached samples from those bumpy surfaces. Therefore, a simple method is needed to obtain detached samples directly from the tissue surface. If the lesion site can be further accurately located, the surgeon can remove the lesion for the patient as soon as possible. Summary of the Invention

[0005] In order to solve the above problems existing in the prior art, the present invention provides a method for searching for a target substance that can be detached on the surface of a tissue block, the method comprising the steps of:

[0006] 1) Tightly imprinting a tissue block containing a target substance that can be detached onto the surface of a deformable material and then removing it;

[0007] 2) placing the deformable material having the target substance retained thereon in close contact with an absorptive glass slide;

[0008] 3) Processing the glass slide on which the target substance is adsorbed so that the target substance can be detected or observed.

[0009] Preferably, the target substance is a specific cell and / or cell fragment. In a preferred embodiment of the present invention, the target substance is a tumor cell or a tumor cell with certain characteristics.

[0010] Preferably, in step 1), the tissue block is turned over on the deformable material so that multiple surfaces of the tissue block are tightly impressed on the surface of the deformable material.

[0011] Preferably, the surface area of ​​the tissue block used for stamping is greater than 4 cm 2 , more preferably greater than 8cm 2 .

[0012] Preferably, the surface of the tissue block is uneven.

[0013] Preferably, the deformable material can completely conform to the uneven surface of the tissue block, and will not automatically rebound after the tissue block is removed.

[0014] Preferably, the deformable material is viscous; further preferably, the material used to make it is a hydrophilic colloid, which can be selected from one or more of gelatin, carrageenan, and agar.

[0015] In a preferred embodiment of the present invention, the deformable material is a mixture of gelatin and carrageenan.

[0016] Furthermore, the close contact in step 2) refers to pressing the deformable material with an uneven surface onto the surface of the adsorptive glass slide to make it flat, so that the detached target substance is transferred to the adsorptive glass slide.

[0017] Preferably, the size of the adsorptive slide can cover the entire surface of the tissue block or the surface of all key areas.

[0018] Preferably, the adsorptive glass slide is circular, and its diameter is preferably greater than 3 cm, and more preferably greater than 5 cm or 8 cm.

[0019] Preferably, the treatment in step 3) refers to directly placing the glass slide under a microscope for observation or staining the target substance on the glass slide before placing it under a microscope for observation.

[0020] Furthermore, the method for searching for a target substance that can be shed on the surface of a tissue block further comprises the step of: finding a position on the surface of the tissue block corresponding to the target substance before it sheds.

[0021] In a preferred embodiment of the present invention, after step 1), the position of the key area corresponding to the tissue block is circled on the surface of the deformable material.

[0022] In another preferred embodiment of the present invention, the method for searching for a target substance that can be detached from the surface of a tissue block comprises the following steps:

[0023] 1) Impressing a tissue block containing a target substance that can be detached onto the surface of a deformable material, marking the corresponding positions of the two, and then removing them;

[0024] 2) placing the deformable material having the target substance retained thereon in close contact with an absorptive glass slide, and marking the corresponding position on the absorptive glass slide;

[0025] 3) Processing the glass slide on which the target substance is adsorbed so that the target substance can be detected or observed.

[0026] In another preferred embodiment of the present invention, the position of the target substance before it falls off can be located by setting reference objects on any two or all three of the tissue block, the deformable material, and the absorbent slide.

[0027] Specifically, the method of searching for a target substance that can be detached on the surface of a tissue block according to the present invention may include the following steps:

[0028] 1) tightly imprinting a tissue block containing a target substance that can be detached onto the surface of a deformable material, simultaneously placing a first reference object and a second reference object at corresponding positions on the deformable material and the tissue block, and then separating the two;

[0029] 2) placing the deformable material having the target substance retained thereon in close contact with an absorbent glass slide provided with a third reference object, the third reference object being located at a position corresponding to the first reference object;

[0030] 3) treating the glass slide adsorbed with the target substance so that the target substance can be detected or observed;

[0031] 4) marking the position of the target substance to be detected or observed on a transparent soft film having the same structure as the glass slide, the same markings, and a third reference object;

[0032] 5) The soft membrane is adhered to and covers the surface of the tissue block by correspondingly connecting the second reference object and the third reference object.

[0033] Furthermore, the transparent soft membrane can be removed after the original position of the target substance is depicted on the surface of the tissue.

[0034] Preferably, the first reference object, the second reference object and the third reference object are connected with each other by means of a connection structure, such as a sleeve connection, a clip connection or an adhesive connection.

[0035] In a preferred embodiment of the present invention, the connection method is socket connection, the first reference object is a plurality of needle-like structures, the second reference object is a corresponding number of cylindrical structures whose inner diameter is the same as the diameter of the needle-like structure, and the third reference object is a corresponding number of circular hole structures whose inner diameter is the same as the outer diameter of the cylindrical structure, so that the third reference object and the second reference object can be completely matched, and the side wall of the cylindrical structure is relatively thin, not exceeding 0.5 mm, so that the error between the third reference object and the first reference object is minimized, or the inner diameter of the circular hole of the third reference object can also be set to be adjustable.

[0036] The present invention provides a method for searching for detachable target substances from the surface of a tissue block. The method is simple to operate and can be used for rapid pathological diagnosis during surgery. Since the method omits the process of making frozen sections, it only requires the tissue sample to be imprinted on the surface of a deformable material. The obtained detachable substances (such as cells) can be directly used for pathological diagnosis. Therefore, the method can obtain preliminary diagnostic results more quickly. The method is very suitable for tissue samples with uneven surfaces or scattered lesions. Furthermore, the method can directly locate the position of the lesion on the original tissue, allowing the surgeon to perform lesion resection or precise sampling during surgery. The method has good clinical application prospects.

[0037] The concept, specific structure and technical effects of the present invention will be further described below in conjunction with the accompanying drawings to fully understand the purpose, characteristics and effects of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 1 is an operational schematic diagram of step 1) of the method of the present invention;

[0039] Figure 2 This is a schematic diagram of the flipping imprinting trajectory of the tissue block;

[0040] Figure 3 2 is an operational diagram of step 2) of the method of the present invention;

[0041] Figure 4 This is a schematic diagram of the skin tissue structure containing a tumor;

[0042] Figure 5 It is a schematic diagram of the surface structure of the deformable material after imprinting;

[0043] Figure 6 is a schematic diagram of the structure and operation of the tissue block, deformable material and reference object in Example 3;

[0044] Figure 7 is a schematic structural diagram of the glass slide and the transparent soft film in Example 3;

[0045] In the figure: 1-tissue block, 2-deformable material, 3-absorbent slide, 4-epidermis, 5-fat layer, 6-tumor foci, 7-first reference object, 8-second reference object, 9-transparent soft membrane, 10-third reference object. DETAILED DESCRIPTION

[0046] Example 1

[0047] like Figure 1 As shown, a tumor tissue block 1 with a flat or uneven cut surface is tightly pressed on the surface of a deformable material 2 (the deformable material 2 can be made of a mixture of gelatin and carrageenan). Figure 2 As shown, the tissue block is turned over so that each surface is imprinted on the surface of the deformable material 2. At the same time, all surfaces can be marked on the deformable material 2, such as six surfaces marked as a, b, c, d, e, and f. During imprinting, the deformable material will deform along with the convex or concave surfaces of the tissue block. Then, the imprinted surface of the deformable material 2 is placed downward and in close contact with the absorbent slide 3 ( Figure 3 ), the embossed surface of the deformable material 2 is also flattened, allowing the detached target material to completely contact the surface of the glass slide 3 and be adsorbed thereon. The deformable material 2 and the absorptive glass slide 3 can be made into structures of identical shape and size to facilitate transfer by pressure. Finally, the glass slide with the adsorbed detached cells (including tumor cells) can be observed directly under a microscope or after treatment such as fixation and staining.

[0048] Example 2

[0049] For skin tissue containing thick fat, since the fat cutting edge is not smooth and the tumor lesions may be more dispersed, it is more suitable to use the method of the present invention to collect exfoliated cells and thus find tumor cells. Figure 4 The figure shows a piece of skin tissue removed during surgery, which includes the top layer of epidermis 4, a rough and thick fat layer 5, and normal tissue or tumor lesions 6 scattered throughout the fat. The nature of the tumor cannot be directly determined by the naked eye from the cut skin tissue, but it is relatively complicated to take samples from all suspicious areas. Therefore, in this embodiment, the entire piece of skin tissue containing the suspicious area is tightly imprinted on the surface of the deformable material 2. After the imprinting is completed, the surface of the deformable material 2 also forms corresponding protrusions or pits (such as Figure 5 As shown in the figure, the doctor can draw circles and mark some symbols around the key bumps or pits so that after finding the tumor cells, they can correspond to their positions on the skin tissue. The surgeon can also further sample or remove the tumor tissue according to the position indicated by the pathology department.

[0050] Example 3

[0051] Figure 6 and Figure 7 The embodiment of the present invention containing reference objects is shown. Two needle-shaped first reference objects 7 are set on the deformable material 2 and two corresponding second reference objects 8 are set on the deformable material 2. The second reference objects 8 are cylindrical structures with the same inner diameter as the first reference object 7 so that the two are exactly matched. The second reference objects 8 can be removed from the first reference object 7. When the tissue cut 1 is pressed downward onto the deformable material 2, the second reference objects 8 will be inserted into the tissue. When the tissue cut 1 is removed, the second reference objects 8 can remain in the tissue and can be removed. The surface of the deformable material 2 will retain the shape of the contact surface of the tissue cut 1, and the first reference object 7 will also remain on the deformable material 2. Then, the third reference object 10 with a circular hole structure on the adsorptive glass slide 3 is placed on the first reference object 7. The inner diameter of the circular hole structure is the same as the outer diameter of the second reference object 8, and the side wall thickness of the second reference object 8 is less than 0.5 mm, so that the third reference object 10 is basically consistent with the first reference object 7. After turning over the deformable material 2, press the deformable material 2 and the adsorptive glass slide 3 downward, and manually adjust the original This concave portion is pressed against the surface of the absorptive glass slide 3. Due to the absorptive properties of the glass slide 3, cells originally adhered to the deformable material 2 can be transferred to the surface of the absorptive glass slide 3. The absorptive glass slide 3 is then subjected to conventional processing (such as staining). After processing, the specific tumor cells can be found under a microscope and their locations can be determined on the absorptive glass slide 3 (using the lines on the glass slide). This embodiment also provides a transparent soft film 9 that is identical to the absorptive glass slide 3 and is also provided with two third reference objects 10. The doctor can match the tumor cell locations on the absorptive glass slide 3 to the transparent soft film 9 based on the lines on the soft film. The transparent soft film 9 is then placed on the contact surface of the tissue section 1. Since the soft film material is puncturable, the doctor can use a pen or other marking tool to directly mark the tumor cell locations on the transparent soft film 9 on the surface of the tissue section 1 through the soft film. The marked tissue section 1 can be returned to the operating room or photographed and sent to the operating room for reference during further tumor resection or sampling.

[0052] The above describes in detail the preferred embodiments of the present invention. It should be understood that those skilled in the art can make numerous modifications and variations based on the concepts of the present invention without inventive effort. Therefore, any technical solutions that can be derived by those skilled in the art through logical analysis, reasoning, or limited experimentation based on the concepts of the present invention and the prior art should be within the scope of protection defined by the claims.

Claims

1. A method for searching for a target substance that can be detached on the surface of a tissue block, characterized in that: The surface of the tissue block is uneven, and the target substance is specific cells and / or cell fragments, comprising the steps of: 1) Tightly imprinting a tissue block containing a target substance that can be detached onto the surface of a deformable material and then removing it; 2) placing the deformable material having the target substance retained thereon in close contact with an adsorptive glass slide; 3) treating the glass slide adsorbed with the target substance so that the target substance can be detected or observed; in step 1), the tissue block is flipped over on the deformable material so that multiple surfaces of the tissue block are tightly impressed on the surface of the deformable material; The treatment described in step 3) refers to directly placing the glass slide under a microscope for observation or staining the target substance on the glass slide before placing it under a microscope for observation.

2. The method for searching for a target substance that can be detached from the surface of a tissue block according to claim 1, wherein: The method further comprises the step of finding the position on the surface of the tissue block corresponding to the target substance before it falls off.

3. The method for searching for a target substance that can be detached from the surface of a tissue block according to claim 2, wherein: After step 1), the position of the key area of ​​the corresponding tissue block is circled on the surface of the deformable material.

4. The method for searching for a target substance that can be detached from the surface of a tissue block according to claim 2, wherein: The position of the target substance before it falls off is located by setting reference objects on any two or all three of the tissue block, the deformable material, and the absorbent slide.

5. The method for searching for a target substance that can be detached from the surface of a tissue block according to claim 1, wherein: Step 1) further includes: simultaneously marking the corresponding positions of the tissue block and the surface of the deformable material and then moving them away; step 2) further includes simultaneously marking the corresponding positions on the adsorptive glass slide.

6. The method for searching for a target substance that can be detached from the surface of a tissue block according to claim 1, wherein: Step 1) further includes: simultaneously placing a first reference object and a second reference object at corresponding positions on the tissue block and the deformable material, and then separating the two; step 2) further includes placing a third reference object on the adsorptive slide, the third reference object corresponding to the position of the second reference object; In addition, the method further comprises the steps of: 4) marking the position of the target substance to be detected or observed on a transparent soft film having the same structure as the glass slide, the same markings, and a third reference object; 5) The soft membrane is adhered to and covers the surface of the tissue block by correspondingly connecting the first reference object and the third reference object.

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