CN109975090B - Preparation method of thyroid and mammary gland fine needle punctured cell tissue block - Google Patents
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/36—Embedding or analogous mounting of samples
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Abstract
The invention provides a preparation method of a cell tissue block punctured by a fine needle of thyroid and mammary gland. The method comprises the following steps: a. centrifuging non-infectious pleural fluid or ascites in advance to obtain pleural fluid or ascites supernatant; b. injecting the pleural fluid or ascites supernatant into a centrifuge tube; c. injecting the thyroid or breast specimen punctured by the fine needle into a centrifuge tube, and immediately injecting 95% ethanol into the centrifuge tube; d. centrifuging the centrifuge tube, removing the supernatant to obtain cell precipitate, adding 4% neutral buffer formaldehyde fixing solution, and performing conventional embedding by adopting a paraffin tissue specimen processing procedure after the cell precipitate in the centrifuge tube is solidified. The preparation method of the invention can ensure that tissue fragments and free-floating single cells are not lost, the shape and structure of cytoplasm are well preserved, and the cytoplasm can be continuously or repeatedly sliced, and is suitable for various stains, thereby improving the diagnosis effect.
Description
Technical Field
The invention relates to the technical field of preparation of cell tissue blocks, in particular to a preparation method of a thyroid and mammary gland fine needle puncture cell tissue block.
Background
The incidence of thyroid cancer and breast cancer is rapidly increasing year by year in the world, wherein the incidence of thyroid cancer is increased year by year at a rate of 6%. At present, the average incidence rate of thyroid cancer in China is 7.7/10 ten thousand, and the incidence rate of malignant tumors ranks the fifth place. 85% of Thyroid adenocarcinomas are Papillary carcinomas (PTC). One particular biological behavior of PTC is the early development of regional lymph node metastasis. Although differentiated thyroid cancer heals well, during the last 30 years of follow-up, about 30% of patients with thyroid cancer will relapse, and eventually a small percentage of patients will die from thyroid cancer. The preoperative diagnosis of thyroid cancer is mainly a thyroid puncture technology.
Thyroid gland puncture is a simple and feasible method which can achieve pathological diagnosis to a certain extent. The puncture is divided into fine and coarse needles. The diameter of the fine needle head is 21-26G, and the diameter of the coarse needle puncture needle head is 16-18G. The most different point of the fine needle puncture is that the sample amount is small, so the fine needle puncture is mostly used for preparing a cell smear, most or all of the histological structures and intercellular matrixes in the sample are lost, and the typing of tumors is not accurate enough, so the limitation of the fine needle aspiration cytology is caused. Secondly, the cell smear can only be directly subjected to immunohistochemical staining without antigen retrieval, but the background is slightly colored, target cells in the smear are scattered and distributed, the number of the target cells is small, the target cells are easily damaged in the smear making process, and the detection result is unstable due to the problems of limitation of the number of smear slides, reagent waste, incapability of marking various antibodies on the cell smear, incapability of correctly judging the staining result and the like.
There are many documents on the conventional methods for producing cell masses, and they can be roughly classified into direct methods and indirect methods, i.e., production methods using a matrix material. The indirect method comprises the following steps: agar cell block preparation, thrombin-plasma cell block preparation, Histogel preparation, etc. The method has less cell loss, and the matrix material can net the cells in a 'tangible' structure under corresponding conditions, thereby being beneficial to material taking and embedding. However, agar and HistoGel have high density after solidification, and the liquid inside and outside the tissue is not easy to permeate, so the steps of fixing, dehydrating, wax penetration and the like are required for a longer time, and the method needs to be groped and has complex operation. The thrombin-plasma entrapment may affect the results of experiments such as IHC or DNA extraction.
At present, due to the particularity of cells such as thyroid gland and mammary gland, the technology for preparing cell tissue blocks by using fine needle puncture specimens is not mature, so that the technology cannot be popularized and applied. The practice type puncture needle (diameter 0.8 mm) is adopted by the remnant of friendship hospital, and the new protein ethanol coagulation-formaldehyde fixation method is adopted to make thyroid cell tissue block, the puncture needle is relatively thick, and the specimen is placed on the glass sheet, so that the cells are extruded and damaged in the process of gathering, the structures of cell membrane and cell nucleus are not clear, and the diagnosis and further immunohistochemical staining effects are influenced. In conclusion, finding a better method for preparing thyroid and mammary gland cell tissue blocks by fine needle puncture has important significance for relieving the pain of the puncture of a patient and improving the accuracy and efficiency of diagnosis.
Disclosure of Invention
The invention aims to provide a preparation method of thyroid and mammary gland fine needle puncture cell tissue blocks, and solves the problem that thyroid and mammary gland specimens and cell smears which are not suitable for fine needle puncture in the existing tissue block preparation method are difficult to accurately diagnose.
The technical scheme adopted by the invention for realizing the purpose is as follows: a preparation method of thyroid and mammary gland fine needle puncture cell tissue blocks comprises the following steps:
a. centrifuging non-infectious pleural fluid or ascites in advance to obtain pleural fluid or ascites supernatant;
b. cutting a disposable 10mL centrifuge tube by half, and injecting 0.5-2 mL of hydrothorax or ascites supernatant into the centrifuge tube;
c. injecting the thyroid or breast specimen punctured by the fine needle into a centrifuge tube, immediately agglutinating the punctured tissue in pleural effusion or ascites supernatant into a strip shape or a block shape, and immediately injecting 0.5-2 mL of 95% ethanol into the centrifuge tube;
d. centrifuging the centrifuge tube and removing the supernatant to obtain cell sediment, adding 4% neutral buffer formaldehyde fixing solution, taking out the cell sediment in the centrifuge tube after fixing for 3-5 h, putting the cell sediment into embedding paper, then putting the embedding paper into a dehydration embedding box, and performing conventional embedding by adopting a paraffin tissue specimen processing procedure.
In the step a, the obtained pleural effusion or ascites supernatant is pre-checked, and the method comprises the following steps: injecting 0.5-2 mL of pleural effusion or ascites supernatant into a centrifuge tube, adding 95% ethanol with the same volume, centrifuging, removing supernatant to obtain a precipitate, adding 5-10 times of 4% neutral buffer formaldehyde fixing solution, taking out the precipitate after solidification, putting the precipitate into embedding paper, then putting the embedding paper into a dehydration embedding box, performing conventional embedding by adopting a paraffin tissue specimen processing procedure, and performing section and HE staining and then sealing microscopic examination to ensure that no cell component is detected by microscopic examination.
In the step c, the needle head for fine needle puncture is 23G and 0.573mm in diameter.
In step c, the volume of 95% ethanol used is the same as the volume of pleural fluid or ascites supernatant.
And c, flushing the needle head by using pleural effusion or ascites, and then injecting the puncture tissue in the needle head into the centrifugal tube.
In the step d, centrifuging for 10 min under the centrifugal condition of 2000 r/min; the volume of the added 4% neutral buffered formaldehyde fixing solution is 5-10 times of the volume of the cell sediment obtained by centrifugation.
In the thyroid fine needle puncture sample collected by the invention, even if the cell amount of the sample is small or the cells are loose, tissue fragments and free floating single cells can be ensured to be free from loss after the protein screening of the hydrothorax and ascites, the morphology and the structure of cytoplasm are well preserved, and the nuclear sulcus and the nucleolus are also clear, thereby improving the diagnosis effect. Meanwhile, the antibody with the stained immunocytochemistry cell membrane or cytoplasm can be interpreted; can be used for gene detection; and the hydrothorax and ascites pathology department has the advantages of daily use, no cost, convenience, easy obtaining and no ethical problems such as patient privacy.
The cell tissue block manufacturing technology organically combines the thyroid gland and mammary gland puncture cytology with the fixing and processing of the specimen in the histology and the paraffin embedding technology, not only preserves the good cell morphological characteristics of the puncture tissue, but also can further explore the tissue morphological characteristics, is convenient for the further research of the auxiliary detection of immunocytochemistry staining and the like, and the practice proves that the application effect is good, the detection cost of the BRAFV600E can be saved, and certain economic and social benefits are formed. The cell block technology can be effectively applied to daily cytology work and is also suitable for routine development in hospitals at all levels.
The invention takes the pleural effusion as a carrier, the cell tissue block can be continuously or repeatedly sliced and stained by using various antibodies (including BRAFV 600E) for immunohistochemistry and the like, and the cell block and the white film can be stored for a long time for later more routine detection, even molecular detection, thereby making up the inherent defect that the cell smear can not be stained for multiple varieties. In addition, the technology can be applied to the preparation of lymph node, lung, pancreas, parotid gland and other system blocks.
Drawings
FIG. 1 is a graphical representation of thyroid cell morphology at different magnifications after HE staining in example 1 of the invention.
FIG. 2 is a morphogram of 20X10 thyroid cells after immunohistochemical staining in example 1 of the present invention, wherein panel (a) is strongly positive for CK19 membrane and panel (b) is strongly positive for TTF-1 nucleus.
FIG. 3 is a morphogram of thyroid cells after HE staining in comparative example 1.
FIG. 4 is a morphogram of breast cells at different magnifications after HE staining in example 2 of the invention.
Detailed Description
The production process of the present invention is described in detail below with specific examples.
Example 1: and (3) preparing a thyroid fine needle puncture cell tissue block.
Firstly, a pre-experiment step: taking non-infectious pleural effusion, centrifuging twice to obtain 100mL of supernatant, respectively taking 0.5mL, 1mL, 1.5mL and 2mL of supernatant, respectively injecting the supernatants into a disposable 10mL centrifuge tube, adding 95% ethanol with the same volume, gently mixing, centrifuging at 2000 r/min for 10 min, observing the volume of a precipitated block, taking a block (the volume is about 1.0cm 0.3 cm) with the height of about 0.3cm at the bottom of the centrifuge tube as a sample for standby, and discarding the blocks of other centrifuge tubes.
And (3) discarding the supernatant of the selected sample, and adding 5-10 times of 4% neutral buffer formaldehyde fixing solution. After solidification, the cell sediment solidified in the centrifuge tube is taken out and put into embedding paper, and then put into a dehydration embedding box, and the normal embedding, slicing, HE staining and sealing microscopic examination are carried out by adopting a paraffin tissue specimen processing program (60 ℃ 4% neutral buffered formaldehyde is fixed for 90 min multiplied by 2 times, 60 ℃ absolute ethanol is fixed for 60 min multiplied by 5 times, 60 ℃ xylene is fixed for 60 min multiplied by 3 times, and 60 ℃ paraffin is fixed for 50 min multiplied by 4 times). Under the lens, a large amount of staining substances are seen, and no cell components exist.
II, an experiment step: a disposable 10mL centrifuge tube was cut in half and 0.5mL of the breast water supernatant was injected under microscopic examination (pre-experiments resulted in clumps approximately 0.3cm in height). Washing the 23G thyroid gland puncture needle head punctured by the fine needle with hydrothorax or ascites, then injecting the puncture specimen in the needle head into the centrifugal tube, and immediately agglutinating the puncture tissue into a strip shape or a block shape.
And (2) immediately injecting 0.5mL of 95% ethanol into a centrifugal tube, centrifuging the centrifugal tube at 2000 r/min for 5 min, removing the supernatant, adding 5-10 times of 4% neutral buffer formaldehyde fixing solution, solidifying cell precipitates in the centrifugal tube after 3h-5h, taking out, placing the cell precipitates in embedding paper, then placing the embedding paper into a dehydration embedding box, performing conventional embedding, slicing, HE dyeing and the like by adopting a paraffin tissue specimen processing procedure, and sealing and performing microscopic examination. The tissue block microscopic examination results are shown in fig. 1 and 2. As can be seen from the HE slice, 2x10 has abundant cells, and the number of the cells can reach more than ten thousand; at 4x10, papillary structures and follicular segments can be seen, with the number of cells in follicular segments greater than 10; 10x10, clear and crowded cell nuclei; 20 × 10, the cytoplasm was well preserved and the nuclear groove was visible; in 40 × 10, the nuclear groove is clear, the nuclear membrane is thickened, and 1-2 nucleoli can be seen; at 100x10, the nucleolus is clearer, the nucleoli is clear, and the cytoplasm is kept intact in an oil lens.
Comparative example 1: the plasma-thrombin method produces a thyroid cell tissue mass.
Taking a 10mL centrifuge tube, adding 4% neutral buffered formaldehyde solution, injecting a thyroid 23G needle puncture sample into the centrifuge tube, centrifuging for 5 min at 2000 r/min, discarding the supernatant, adding a plurality of drops of fresh human plasma suspension cell sediment, adding a plurality of drops of thrombin, uniformly mixing, coagulating into blocks, wrapping the sediment with a piece of mirror paper, placing the wrapped sediment into an embedding box, fixing in 10% neutral formaldehyde solution for 2h, performing conventional embedding dehydration, and performing microscopic examination, wherein the result is shown in fig. 3. As can be seen from FIG. 3, blood coagulation, low cellular content, unclear nuclear and cytoplasmic structures, indicate that it is difficult to ensure a satisfactory cell mass with the plasmatic-thrombin method because the clot formed by the combination of cells with plasma and thrombin is generally sparse and loose, easily detached on H & E slices, and the cell mass is very low.
Example 2: preparing a cell tissue block punctured by a fine mammary needle.
Firstly, a pre-experiment step: taking non-infectious ascites to obtain 100mL of supernatant after twice centrifugation, respectively taking 0.5mL, 1mL, 1.5mL and 2mL of supernatant to respectively inject into a disposable 10mL centrifuge tube, adding 95% ethanol with the same volume, gently mixing, centrifuging for 5 min at 2000 r/min, observing the volume of a precipitated block, taking a block (the volume is about 1.0cm x 0.3 cm) with the height of about 0.3cm at the bottom of the centrifuge tube as a sample for later use, and discarding the blocks of other centrifuge tubes.
And (3) discarding the supernatant of the selected sample, and adding 5-10 times of 4% neutral buffer formaldehyde fixing solution. After solidification, taking out the cell sediment solidified in the centrifuge tube, putting the cell sediment into embedding paper, then putting the cell sediment into a dehydration embedding box, carrying out routine embedding, slicing and HE staining by adopting a paraffin tissue specimen processing procedure, and sealing and carrying out microscopic examination. Under the lens, a large amount of staining substances are seen, and no cell components exist.
II, an experiment step: a disposable 10mL centrifuge tube was cut in half and 0.5mL of the microscopically examined ascites supernatant was injected (a block of approximately 0.3cm height could be formed in the preliminary experiment). The 23G mammary gland puncture needle head punctured by the fine needle is washed by pleural effusion or ascites, then a puncture specimen in the needle head is injected into a centrifugal tube, and the puncture tissue is immediately aggregated into a strip shape or a block shape.
And (2) immediately injecting 0.5mL of 95% ethanol into a centrifugal tube, centrifuging the centrifugal tube at 2000 r/min for 10 min, removing the supernatant, adding 5-10 times of 4% neutral buffer formaldehyde fixing solution, solidifying cell precipitates in the centrifugal tube after 3h-5h, taking out, placing the cell precipitates in embedding paper, then placing the embedding paper into a dehydration embedding box, performing conventional embedding, slicing, HE dyeing and the like by adopting a paraffin tissue specimen processing procedure, and sealing and performing microscopic examination. The tissue block microscopic examination results are shown in fig. 4. It can be seen from FIG. 4 that the cells were abundant and the nuclei and cytoplasm were well preserved.
Claims (5)
1. A preparation method of thyroid and mammary gland fine needle puncture cell tissue blocks is characterized by comprising the following steps:
a. centrifuging non-infectious pleural fluid or ascites in advance to obtain pleural fluid or ascites supernatant; the obtained pleural fluid or ascites supernatant is pre-checked, and the method comprises the following steps: injecting 0.5-2 mL of pleural effusion or ascites supernatant into a centrifuge tube, adding 95% ethanol with the same volume, centrifuging, removing supernatant to obtain a precipitate, adding 4% neutral buffer formaldehyde fixing solution with the volume 5-10 times that of the precipitate, taking out the precipitate after solidification, putting the precipitate into embedding paper, then putting the embedding paper into a dehydration embedding box, performing conventional embedding by adopting a paraffin tissue specimen processing procedure, and performing section and HE staining and then sealing microscopic examination to ensure no cell component in microscopic examination;
b. cutting a disposable 10mL centrifuge tube by half, and injecting 0.5-2 mL of hydrothorax or ascites supernatant into the centrifuge tube;
c. injecting the thyroid or breast specimen punctured by the fine needle into a centrifuge tube, immediately agglutinating the punctured tissue in pleural effusion or ascites supernatant into a strip shape or a block shape, and immediately injecting 0.5-2 mL of 95% ethanol into the centrifuge tube;
d. centrifuging the centrifuge tube and removing the supernatant to obtain cell sediment, adding 4% neutral buffer formaldehyde fixing solution, taking out the cell sediment in the centrifuge tube after fixing for 3-5 h, putting the cell sediment into embedding paper, then putting the embedding paper into a dehydration embedding box, and performing conventional embedding by adopting a paraffin tissue specimen processing procedure.
2. The method for preparing a thyroid and breast fine needle punctured cell tissue mass as claimed in claim 1, wherein in step c, the needle used for fine needle puncture is 23G and 0.573mm in diameter.
3. The method of claim 1, wherein the volume of 95% ethanol used in step c is the same as the volume of pleural fluid or ascites supernatant.
4. The method for preparing a thyroid and breast fine needle punctured cell tissue mass according to claim 1, wherein in the step c, the needle is washed with pleural fluid or ascites, and then the punctured tissue in the needle is injected into the centrifuge tube.
5. The method for preparing a thyroid and breast fine needle punctured cell tissue mass according to claim 1, wherein in the step d, the centrifugation condition is 2000 r/min for 10 min; the volume of the added 4% neutral buffered formaldehyde fixing solution is 5-10 times of the volume of the cell sediment obtained by centrifugation.
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| CN114441265B (en) * | 2022-01-20 | 2023-09-12 | 南京市中医院 | Preparation method of cell wax block and application of cell wax block in thyrocentesis smear |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0856729A2 (en) * | 1997-01-30 | 1998-08-05 | Shandon Scientific Limited | Tissue processing apparatus and method |
| CN102680295A (en) * | 2012-05-21 | 2012-09-19 | 江苏省中医院 | Method for extracting residual nucleic acid from HE (haematoxylin eosin) dyeing piece |
| EP2542883A1 (en) * | 2010-03-04 | 2013-01-09 | Ventana Medical Systems, Inc. | Processing system for processing specimens using acoustic energy |
| CN105606413A (en) * | 2015-10-16 | 2016-05-25 | 中南大学 | Preparation method of renal biopsy electron microscopy sample |
| CN106769278A (en) * | 2016-11-18 | 2017-05-31 | 张震 | A kind of cell block reagent preparation box and preparation method thereof |
| CN108020452A (en) * | 2017-12-14 | 2018-05-11 | 漳州卫生职业学院 | Cell block based on courageous and upright Pleural effusions and preparation method thereof |
-
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Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0856729A2 (en) * | 1997-01-30 | 1998-08-05 | Shandon Scientific Limited | Tissue processing apparatus and method |
| EP2542883A1 (en) * | 2010-03-04 | 2013-01-09 | Ventana Medical Systems, Inc. | Processing system for processing specimens using acoustic energy |
| CN102680295A (en) * | 2012-05-21 | 2012-09-19 | 江苏省中医院 | Method for extracting residual nucleic acid from HE (haematoxylin eosin) dyeing piece |
| CN105606413A (en) * | 2015-10-16 | 2016-05-25 | 中南大学 | Preparation method of renal biopsy electron microscopy sample |
| CN106769278A (en) * | 2016-11-18 | 2017-05-31 | 张震 | A kind of cell block reagent preparation box and preparation method thereof |
| CN108020452A (en) * | 2017-12-14 | 2018-05-11 | 漳州卫生职业学院 | Cell block based on courageous and upright Pleural effusions and preparation method thereof |
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