CN109975090A - A kind of preparation method of thyroid gland, mammary gland Fine-needle puncture cell tissue block - Google Patents
A kind of preparation method of thyroid gland, mammary gland Fine-needle puncture cell tissue block Download PDFInfo
- Publication number
- CN109975090A CN109975090A CN201910297308.XA CN201910297308A CN109975090A CN 109975090 A CN109975090 A CN 109975090A CN 201910297308 A CN201910297308 A CN 201910297308A CN 109975090 A CN109975090 A CN 109975090A
- Authority
- CN
- China
- Prior art keywords
- centrifuge tube
- hydrothorax
- fine
- preparation
- needle puncture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 210000001519 tissue Anatomy 0.000 title claims abstract description 33
- 210000001685 thyroid gland Anatomy 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- 210000005075 mammary gland Anatomy 0.000 title claims abstract description 18
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 29
- 239000006228 supernatant Substances 0.000 claims abstract description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 206010003445 Ascites Diseases 0.000 claims abstract description 23
- 206010048612 Hydrothorax Diseases 0.000 claims abstract description 23
- 230000007935 neutral effect Effects 0.000 claims abstract description 13
- 239000012188 paraffin wax Substances 0.000 claims abstract description 11
- 239000002244 precipitate Substances 0.000 claims abstract description 11
- 238000004043 dyeing Methods 0.000 claims abstract description 9
- 230000008520 organization Effects 0.000 claims abstract description 9
- 238000002347 injection Methods 0.000 claims abstract description 7
- 239000007924 injection Substances 0.000 claims abstract description 7
- 238000007711 solidification Methods 0.000 claims abstract description 7
- 230000008023 solidification Effects 0.000 claims abstract description 7
- 208000015181 infectious disease Diseases 0.000 claims abstract description 3
- 230000002458 infectious effect Effects 0.000 claims abstract description 3
- 238000000034 method Methods 0.000 claims description 14
- 235000019441 ethanol Nutrition 0.000 claims description 11
- 238000000386 microscopy Methods 0.000 claims description 11
- 238000005119 centrifugation Methods 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 5
- 239000013049 sediment Substances 0.000 claims description 5
- 239000004615 ingredient Substances 0.000 claims description 4
- 230000001376 precipitating effect Effects 0.000 claims description 4
- 239000000047 product Substances 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 48
- 210000000805 cytoplasm Anatomy 0.000 abstract description 7
- 238000003745 diagnosis Methods 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 5
- 239000012634 fragment Substances 0.000 abstract description 2
- 229960004756 ethanol Drugs 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 239000008280 blood Substances 0.000 description 6
- 201000002510 thyroid cancer Diseases 0.000 description 6
- 208000024770 Thyroid neoplasm Diseases 0.000 description 5
- 229950003499 fibrin Drugs 0.000 description 5
- 210000004940 nucleus Anatomy 0.000 description 5
- 102000009123 Fibrin Human genes 0.000 description 4
- 108010073385 Fibrin Proteins 0.000 description 4
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 208000002151 Pleural effusion Diseases 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 2
- 208000009453 Thyroid Nodule Diseases 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 230000000762 glandular Effects 0.000 description 2
- 238000011532 immunohistochemical staining Methods 0.000 description 2
- 238000003364 immunohistochemistry Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 208000025962 Crush injury Diseases 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 101710114425 Homeobox protein Nkx-2.1 Proteins 0.000 description 1
- 102100027893 Homeobox protein Nkx-2.1 Human genes 0.000 description 1
- 101000998011 Homo sapiens Keratin, type I cytoskeletal 19 Proteins 0.000 description 1
- 102100033420 Keratin, type I cytoskeletal 19 Human genes 0.000 description 1
- 208000007433 Lymphatic Metastasis Diseases 0.000 description 1
- 206010033701 Papillary thyroid cancer Diseases 0.000 description 1
- 101710088547 Thyroid transcription factor 1 Proteins 0.000 description 1
- 101710159262 Transcription termination factor 1 Proteins 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000006727 cell loss Effects 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 208000015799 differentiated thyroid carcinoma Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000005357 flat glass Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 238000003365 immunocytochemistry Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000000116 mitigating effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000002445 nipple Anatomy 0.000 description 1
- 210000000633 nuclear envelope Anatomy 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 201000010198 papillary carcinoma Diseases 0.000 description 1
- 210000003681 parotid gland Anatomy 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 238000010882 preoperative diagnosis Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000008719 thickening Effects 0.000 description 1
- 208000030829 thyroid gland adenocarcinoma Diseases 0.000 description 1
- 208000030901 thyroid gland follicular carcinoma Diseases 0.000 description 1
- 208000030045 thyroid gland papillary carcinoma Diseases 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
Classifications
-
- 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Sampling And Sample Adjustment (AREA)
Abstract
The present invention provides the preparation methods of a kind of thyroid gland, mammary gland Fine-needle puncture cell tissue block.The following steps are included: a, being in advance centrifuged non-infectious hydrothorax or ascites, hydrothorax or ascites supernatant are obtained；B, hydrothorax or ascites supernatant is taken to be injected into centrifuge tube；C, the thyroid gland of fine needle puncture or mammary gland sample are injected into centrifuge tube, are taken in 95% ethanol injection centrifuge tube immediately after；D, centrifuge tube is centrifuged to and is abandoned supernatant, obtains cell precipitate, 4% neutral buffered formalin fixer is then added, after the cell precipitate solidification in centrifuge tube, conventional embedding is carried out using paraffin organization sample disposal program.The unicellular no loss that preparation method of the invention can guarantee fragment of tissue and freely float, cytoplasm form and structure save well, can continuously or be repeatedly sliced, and are suitable for a variety of dyeing, to improve diagnosis effect.
Description
Technical field
The present invention relates to cell tissue block preparation technical field, a kind of thyroid gland, mammary gland fine needle puncture are related in particular to
The preparation method of cell tissue block.
Background technique
The disease incidence of thyroid cancer, breast cancer worldwide is in the trend rapidly risen year by year, wherein thyroid cancer
Disease incidence is every year with 6% speed cumulative year after year.Average thyroid cancer incidence domestic at present is 7.7/10 ten thousand, in malignant tumour
The 5th is occupied in disease incidence ranking.85% thyroid adenocarcinoma be papillary carcinoma (Papillary Thyroid Carcinoma,
PTC).The biological behaviour that one kind of PTC is special is early stage generation area lymphatic metastasis.Although differentiated thyroid carcinoma more after
Very well, but during nearly follow-up in 30 years, about 30% thyroid cancer patients can recur, and final subset of patients can die of first
Shape gland cancer.Preoperative diagnosis thyroid cancer is mainly thyroid needle technology.
Thyroid needle is a kind of method that is easy, easy and can reaching pathological diagnosis to a certain extent.It punctures
It is divided into fine needle and bodkin.Fine needle needle diameter 21-26G, coarse needle bioptic needle diameter 16-18G.Fine needle puncture, it is maximum not
It is that specimen amount is few, thus is mostly used to prepare cell smear with putting, histological structure and intercellular matrix are most of or complete in sample
Full missing, not accurate enough to the parting of tumour, this results in the maximum limitation of thin needle absorbing.Secondly, cell smear is only
It can be carried out direct immunohistochemical staining, be not necessarily to antigen retrieval, but background coloration is shallower, target cell is dispersed in smear
Distribution, rare numbers, and cytoclasis is be easy to cause during film-making, and there is the limitation for smearing slide quantity, examination
Agent waste, cell smear not can be carried out multiple types antibody label and can not correct interpretation coloration result the problems such as, cause to examine
Survey unstable result.
The existing document about cell block preparation method has very much, can generally be divided into direct method and indirect method,
Connection is to have used the preparation method of host material.Indirect method includes following several: agar cell block the preparation method, fibrin ferment-blood
Thick liquid cell block the preparation method and HistoGel the preparation method etc..Above method cell loss is less, and host material under corresponding condition may be used
Cell is enlisted the services of wherein with the structure of " tangible ", be conducive to draw materials and is embedded.But density is larger after agar, HistoGel solidification,
Organize inside and outside liquid to be not easy to permeate, thus to fixation, dehydration, saturating wax and etc. desired time it is longer, need to grope and operate
It is complicated.And fibrin ferment-blood plasma is embedded with and may impact to the result of the experiments such as IHC or DNA extraction.
Currently, cytological samples obtained by fine-needle aspiration makes cell tissue block technology also due to thyroid gland, mammary gland isocellular particularity
It is immature, it causes not promoting and applying.There is small cover etc. more than document report Friendship Hospital to use pen-holding type puncture needle (diameter
0.8mm), thyroid cell tissue block is made using new albumen alcohol solidification-formaldehyde fixation, puncture needle used is relatively
Slightly, and since sample is placed on sheet glass, cell is caused to have crush injury, cell membrane and nuclear structures in the process assembled
It is unintelligible, influence diagnosis and further immunohistochemistry coloring effect.In summary, seek a kind of preferable fine needle puncture preparation first
Shape gland, the method for mammary glandular cell tissue block is painful for mitigation patient's puncture, improves the accuracy and efficiency of diagnosis with important
Meaning.
Summary of the invention
The purpose of the present invention is intended to provide the preparation method of a kind of thyroid gland, mammary gland Fine-needle puncture cell tissue block, with solution
Certainly existing tissue block preparation method is not suitable for the thyroid gland of fine needle puncture, mammary gland sample and cell smear and is difficult to Accurate Diagnosis
Problem.
The present invention is that technical solution used by realizing object above is as follows: a kind of thyroid gland, mammary gland Fine-needle puncture cell
The preparation method of tissue block, comprising the following steps:
A, non-infectious hydrothorax or ascites are centrifuged in advance, obtain hydrothorax or ascites supernatant；
B, it takes disposable 10mL centrifuge tube to cut short half, 0.5 ~ 2mL hydrothorax or ascites supernatant is taken to be injected into centrifuge tube；
C, the thyroid gland of fine needle puncture or mammary gland sample are injected into centrifuge tube, puncturing tissue is in hydrothorax or ascites supernatant
It is aggregated stripped or blocky, takes in the ethanol injection centrifuge tube of 0.5 ~ 2mL 95% immediately after immediately；
D, centrifuge tube is centrifuged to and is abandoned supernatant, obtains cell precipitate, 4% neutral buffered formalin fixer is then added, wait be centrifuged
After cell precipitate in pipe fixes 3 ~ 5h, taking-up is put into embedding paper, is subsequently placed in embedding box for dehydrating, using paraffin organization mark
Present treatment program carries out conventional embedding.
In step a, preliminary examination, method are as follows: by 0.5 ~ 2mL hydrothorax or ascites are carried out to obtained hydrothorax or ascites supernatant
Supernatant is injected into centrifuge tube, and 95% ethyl alcohol of same volume is then added, abandons supernatant after centrifugation, obtains sediment, it is added 5 ~
4% neutral buffered formalin fixer of 10 times of sediment volumes, after object solidification to be precipitated, taking-up is put into embedding paper, is subsequently placed in
In embedding box for dehydrating, using the conventional embedding of paraffin organization sample disposal program progress, sealing microscopy after sliced, HE dyeing, really
Protect the cell-free ingredient of microscopy.
In step c, syringe needle used in the fine needle puncture is 23G, diameter 0.573mm.
In step c, the volume of used 95% ethyl alcohol is identical as the volume of hydrothorax or ascites supernatant.
It, then will be in the puncturing tissue injection centrifuge tube in syringe needle with hydrothorax or ascites Syringe needle Irrigator in step c.
In step d, centrifugal condition is that 2000 r/min are centrifuged 10 min；4% neutral buffered formalin fixer being added
Volume is 5 ~ 10 times of centrifugation gained cell precipitate volume.
In the fine-needle aspiration of thyroid nodules sample that the present invention collects, after the albumen of Pleural effusions is enlisted the services of, even if sample is thin
Born of the same parents' amount less or cell is loose, fragment of tissue and what is freely floated unicellular also can guarantee no loss, cytoplasm morphology and knot
Structure saves well, and core ditch and kernel are also apparent, to improve diagnosis effect.Simultaneously using immunocytochemistry cell membrane or carefully
The antibody of cytoplasm coloring, can interpretation；The detection in terms of gene can be done；And Pleural effusions pathology department can all have daily, not have
Cost is conveniently easy to get, in terms of not also being involved in the problems, such as the ethics such as patient privacy.
Cell tissue block manufacturing technology of the invention consolidates thyroid gland, breast puncture cytology and sample in histology
Fixed, processing, paraffin embedding techniques organically combine, and have not only saved the good cell morphology characteristic of puncturing tissue, but also can be further
Tectology feature is excavated, while the further research for being convenient for the detection of the auxiliary such as immunocytochemical stain is used, it is real
Trampling proves that its application effect is good, and can save thyroid gland BRAFV600E testing cost, forms certain economy and society's effect
Benefit.This cell block technology can effectively apply in daily cytology work, also be suitable for routinely carrying out in situation of all-level hospitals.
The present invention using Pleural effusions as carrier, cell tissue block can by continuously or repeatedly slice using Multiple Antibodies (including
BRAFV600E the dyeing such as immunohistochemistry) is carried out, and cell block and white glue piece can be with long-term preservations in case carry out more in the future
Detection in terms of more conventional detections or even molecule, this, which is just compensated for, not can be carried out multiple multiple types dyeing to cell smear
Inadequate natural endowment.In addition, technique can be applied to the production of the other systems mass such as lymph node, lung, pancreas, parotid gland.
Detailed description of the invention
Fig. 1 is the thyroid cell aspect graph in the embodiment of the present invention 1 after HE is dyed under different amplification.
Fig. 2 is the thyroid cell aspect graph of the 20x10 after immunohistochemical staining in the embodiment of the present invention 1, wherein figure
It (a) is CK19 film strong positive, figure (b) is TTF-1 core strong positive.
Fig. 3 is the thyroid cell aspect graph in comparative example 1 after HE is dyed.
Fig. 4 is the mammary glandular cell aspect graph in the embodiment of the present invention 2 after HE is dyed under different amplification.
Specific embodiment
Below with specific embodiment detailed description of the present invention preparation method.
Embodiment 1: the preparation of fine-needle aspiration of thyroid nodules cell tissue block.
One, preliminary experiment step: negated infectiousness hydrothorax obtains supernatant 100mL after being centrifuged twice, take respectively 0.5mL,
1mL, 1.5mL, 2mL supernatant respectively inject in disposable 10mL centrifuge tube, and 95% ethyl alcohol of equal volume is added, mixes gently,
10 min are centrifuged with 2000 r/min, observe the Mass volum size of precipitating, take the packet of the height about 0.3cm of centrifugation bottom of the tube
Block (volume is about 1.0cm*1.0cm*0.3cm) is spare as sample, and the mass discarding of other centrifuge tubes does not have to.
The sample of selection is abandoned into supernatant, adds 5 ~ 10 times of 4% neutral buffered formalin fixer.It, will be in centrifuge tube after to be solidified
The cell precipitate taking-up of solidification is put into embedding paper, is subsequently placed into embedding box for dehydrating, using paraffin organization sample disposal program
(60 DEG C of 4% neutral buffered formalin fixes 90 min × 2 time, 60 DEG C of 60 min × 5 time of dehydrated alcohol, and 60 DEG C
60 min × 3 time of dimethylbenzene, 60 DEG C of 50 min × 4 time of paraffin) conventional embedding, slice, HE dyeing are carried out, and seal
Gu microscopy.A large amount of powder dye substances, cell-free ingredient are seen under mirror.
Two, experimental procedure: taking disposable 10mL centrifuge tube to cut off half, injects the hydrothorax supernatant 0.5mL through microscopy
(mass that preliminary experiment can form height about 0.3cm).The 23G thyroid needle syringe needle hydrothorax or ascites of fine needle puncture are rushed
It washes, then by the puncture sample injection centrifuge tube in syringe needle, puncturing tissue is aggregated stripped or blocky immediately.
It takes 0.5mL95% ethyl alcohol to inject in centrifuge tube at once, centrifuge tube is centrifuged 5 min with 2000 r/min, in abandoning
Clearly, add 5 ~ 10 times of 4% neutral buffered formalin fixer, centrifuge tube inner cell sediment after 3h-5h, taking-up is put into embedding
Paper is subsequently placed into embedding box for dehydrating, carries out conventional embedding, slice, HE dyeing etc. using paraffin organization sample disposal program, and
Sealing microscopy.Tissue block microscopic examination result is as depicted in figs. 1 and 2.In 2x10 it can be seen from HE slice, cell is very rich, number
Mesh is up to up to ten thousand or more；In 4x10, it can be seen that the cell number of mamilla and folliculus segment, folliculus segment is greater than 10
It is a；In 10x10, nucleus is transparent, crowded；In 20x10, cytoplasm is intact, and nucleus ditch is visible；In 40x10, nucleus
Ditch is clear, thickening of nuclear membrane, it is seen that 1-2 kernel；In 100x10, oil mirror, core ditch is apparent, and kernel understands, cytoplasm saves
Completely.
Comparative example 1: blood plasma-fibrin ferment method prepares thyroid cell tissue block.
10mL centrifuge tube is taken, 4% neutral buffered formalin liquid is added, thyroid gland 23G syringe needle is punctured into sample and injects centrifuge tube
It is interior, 5 min are centrifuged with 2000 r/min, abandon supernatant, then few drops of fresh human plasma suspension cell precipitatings are added thereto,
Be added dropwise again it is blocking to its agglutination after few drops fibrin ferments mix, after taking precipitate is wrapped with lens wiping paper in merging embedded box, in 10%
Property formalin in fix after 2h and carry out conventional dehydration embedding, microscopy, result is as shown in Figure 3.As seen from Figure 3, blood
Solidification, cell component is few, and nucleus and cytoplasm structure are unintelligible, and it is relatively satisfactory thin to illustrate that blood plasma-blood coagulation enzyme process is difficult to ensure
Born of the same parents' amount because the grumeleuse that the combination of cell and blood plasma and fibrin ferment is formed be usually it is sparse, loose, hold on H & E slice
Easy flake, and cell concentration is considerably less.
Embodiment 2: the preparation of mammary gland Fine-needle puncture cell tissue block.
One, preliminary experiment step: negated infectiousness ascites obtains supernatant 100mL after being centrifuged twice, take respectively 0.5mL,
1mL, 1.5mL, 2mL supernatant respectively inject in disposable 10mL centrifuge tube, and 95% ethyl alcohol of equal volume is added, mixes gently,
5 min are centrifuged with 2000 r/min, observe the Mass volum size of precipitating, take the packet of the height about 0.3cm of centrifugation bottom of the tube
Block (volume is about 1.0cm*1.0cm*0.3cm) is spare as sample, and the mass discarding of other centrifuge tubes does not have to.
The sample of selection is abandoned into supernatant, adds 5 ~ 10 times of 4% neutral buffered formalin fixer.It, will be in centrifuge tube after to be solidified
The cell precipitate taking-up of solidification is put into embedding paper, is subsequently placed into embedding box for dehydrating, using paraffin organization sample disposal program,
Conventional embedding, slice, HE dyeing, and sealing microscopy.A large amount of powder dye substances, cell-free ingredient are seen under mirror.
Two, experimental procedure: taking disposable 10mL centrifuge tube to cut off half, injects the ascites supernatant 0.5mL through microscopy
(mass that preliminary experiment can form height about 0.3cm).The 23G breast puncture syringe needle hydrothorax or ascites of fine needle puncture are rinsed,
Then by the puncture sample injection centrifuge tube in syringe needle, puncturing tissue is aggregated stripped or blocky immediately.
It takes 0.5mL95% ethyl alcohol to inject in centrifuge tube at once, centrifuge tube is centrifuged 10 min with 2000 r/min, in abandoning
Clearly, add 5 ~ 10 times of 4% neutral buffered formalin fixer, centrifuge tube inner cell sediment after 3h-5h, taking-up is put into embedding
Paper is subsequently placed into embedding box for dehydrating, carries out conventional embedding, slice, HE dyeing etc. using paraffin organization sample disposal program, and
Sealing microscopy.Tissue block microscopic examination result is as shown in Figure 4.Cell concentration is abundant as seen from Figure 4, and nucleus has been saved with cytoplasm
It is good.
Claims (6)
1. the preparation method of a kind of thyroid gland, mammary gland Fine-needle puncture cell tissue block, characterized in that the following steps are included:
A, non-infectious hydrothorax or ascites are centrifuged in advance, obtain hydrothorax or ascites supernatant；
B, it takes disposable 10mL centrifuge tube to cut short half, 0.5 ~ 2mL hydrothorax or ascites supernatant is taken to be injected into centrifuge tube；
C, the thyroid gland of fine needle puncture or mammary gland sample are injected into centrifuge tube, puncturing tissue is in hydrothorax or ascites supernatant
It is aggregated stripped or blocky, takes in the ethanol injection centrifuge tube of 0.5 ~ 2mL 95% immediately after immediately；
D, centrifuge tube is centrifuged to and is abandoned supernatant, obtains cell precipitate, 4% neutral buffered formalin fixer is then added, wait be centrifuged
After cell precipitate in pipe fixes 3 ~ 5h, taking-up is put into embedding paper, is subsequently placed in embedding box for dehydrating, using paraffin organization mark
Present treatment program carries out conventional embedding.
2. the preparation method of thyroid gland according to claim 1, mammary gland Fine-needle puncture cell tissue block, characterized in that step
In rapid a, preliminary examination, method are carried out to obtained hydrothorax or ascites supernatant are as follows: inject 0.5 ~ 2mL hydrothorax or ascites supernatant
Into centrifuge tube, 95% ethyl alcohol of same volume is then added, abandons supernatant after centrifugation, obtain sediment, 5 ~ 10 times of precipitatings are added
4% neutral buffered formalin fixer of object product, after object solidification to be precipitated, taking-up is put into embedding paper, is subsequently placed in embedding box for dehydrating
In, conventional embedding, sealing microscopy after sliced, HE dyeing, it is ensured that microscopy is without thin are carried out using paraffin organization sample disposal program
Born of the same parents' ingredient.
3. the preparation method of thyroid gland according to claim 1, mammary gland Fine-needle puncture cell tissue block, characterized in that step
In rapid c, syringe needle used in the fine needle puncture is 23G, diameter 0.573mm.
4. the preparation method of thyroid gland according to claim 1, mammary gland Fine-needle puncture cell tissue block, characterized in that step
In rapid c, the volume of used 95% ethyl alcohol is identical as the volume of hydrothorax or ascites supernatant.
5. the preparation method of thyroid gland according to claim 1, mammary gland Fine-needle puncture cell tissue block, characterized in that step
It, then will be in the puncturing tissue injection centrifuge tube in syringe needle with hydrothorax or ascites Syringe needle Irrigator in rapid c.
6. the preparation method of thyroid gland according to claim 1, mammary gland Fine-needle puncture cell tissue block, characterized in that step
In rapid d, centrifugal condition is that 2000 r/min are centrifuged 10 min；The volume for 4% neutral buffered formalin fixer being added is centrifugation
5 ~ 10 times of gained cell precipitate volume.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910297308.XA CN109975090B (en) | 2019-04-15 | 2019-04-15 | Preparation method of thyroid and mammary gland fine needle punctured cell tissue block |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910297308.XA CN109975090B (en) | 2019-04-15 | 2019-04-15 | Preparation method of thyroid and mammary gland fine needle punctured cell tissue block |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109975090A true CN109975090A (en) | 2019-07-05 |
| CN109975090B CN109975090B (en) | 2021-08-10 |
Family
ID=67084430
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910297308.XA Active CN109975090B (en) | 2019-04-15 | 2019-04-15 | Preparation method of thyroid and mammary gland fine needle punctured cell tissue block |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109975090B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112577802A (en) * | 2020-10-15 | 2021-03-30 | 香港大学深圳医院 | Pleural effusion cast-off cell slice staining method |
| CN113074996A (en) * | 2021-03-15 | 2021-07-06 | 陈建华 | Plasma coagulation cell mass method and clinical application thereof |
| CN114441265A (en) * | 2022-01-20 | 2022-05-06 | 南京市中医院 | Preparation method of cell wax block and application of cell wax block in thyroid puncture 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 |
-
2019
- 2019-04-15 CN CN201910297308.XA patent/CN109975090B/en active Active
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 |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112577802A (en) * | 2020-10-15 | 2021-03-30 | 香港大学深圳医院 | Pleural effusion cast-off cell slice staining method |
| CN113074996A (en) * | 2021-03-15 | 2021-07-06 | 陈建华 | Plasma coagulation cell mass method and clinical application thereof |
| CN114441265A (en) * | 2022-01-20 | 2022-05-06 | 南京市中医院 | Preparation method of cell wax block and application of cell wax block in thyroid puncture smear |
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109975090B (en) | 2021-08-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109975090A (en) | A kind of preparation method of thyroid gland, mammary gland Fine-needle puncture cell tissue block | |
| Shidham | CellBlockistry: Chemistry and art of cell-block making–A detailed review of various historical options with recent advances | |
| Nathan et al. | Cell block cytology: improved preparation and its efficacy in diagnostic cytology | |
| Shi et al. | Evaluation of the value of frozen tissue section used as “gold standard” for immunohistochemistry | |
| US8703440B2 (en) | Methods, reagents and instrumentation for preparing impregnated tissue samples suitable for histopathological and molecular studies | |
| Alessandrini et al. | Thyroid-like follicular carcinoma of the kidney: report of two cases with detailed immunohistochemical profile and literature review | |
| CN103940658A (en) | Method for manufacturing paraffin-embedded tissue cell specimen | |
| CN109187146B (en) | Human body cell full-form immunofluorescence staining method and kit | |
| Abele et al. | Smearing techniques for the concentration of particles from fine needle aspiration biopsy | |
| Shi et al. | Self‐clotting method improves cell block preparation | |
| Lv et al. | A novel solution configuration on liquid-based endometrial cytology | |
| CN106536054A (en) | Sample holder for biological samples | |
| Kaneko et al. | A cell‐block preparation using glucomannan extracted from Amorphophallus konjac | |
| Sinkar et al. | Utility of modified ultrafast Papanicolaou stain in cytological diagnosis | |
| Carrara et al. | Histological examination of the diabetic kidney | |
| Rossi et al. | Application of liquid-based preparation to non-gynaecologic exfoliative cytology | |
| CN104487003B (en) | Apparatus and method | |
| CN106769278B (en) | A kind of cell block reagent preparation box and preparation method thereof | |
| Kim et al. | In-house manual construction of high-density and high-quality tissue microarrays by using homemade recipient agarose-paraffin blocks | |
| CN116659987A (en) | Preparation method and application of paraffin section for cytopathology detection | |
| Del Arco et al. | Ber-EP4 staining in effusion cytology: A potential source of false positives | |
| CN107560917B (en) | A kind of processing method of miniature organ | |
| Yu et al. | Occurrence of intercellular spaces (windows) in metastatic adenocarcinoma in serous fluids: a cytomorphologic, histochemical, and ultrastructural study | |
| Shi et al. | Extended application of antigen retrieval technique in immunohistochemistry and in situ hybridization | |
| Lundberg et al. | Immunohistochemical staining of histological fragments derived from salivary gland tumour fine-needle biopsy aspirates |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |