CN108020452A - Cell block based on courageous and upright Pleural effusions and preparation method thereof - Google Patents
Cell block based on courageous and upright Pleural effusions and preparation method thereof Download PDFInfo
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- CN108020452A CN108020452A CN201711342658.0A CN201711342658A CN108020452A CN 108020452 A CN108020452 A CN 108020452A CN 201711342658 A CN201711342658 A CN 201711342658A CN 108020452 A CN108020452 A CN 108020452A
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- 208000002151 Pleural effusion Diseases 0.000 title claims abstract description 40
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 238000001556 precipitation Methods 0.000 claims abstract description 77
- 210000004027 cell Anatomy 0.000 claims abstract description 66
- 210000003743 erythrocyte Anatomy 0.000 claims abstract description 39
- 239000007788 liquid Substances 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 18
- 238000005119 centrifugation Methods 0.000 claims abstract description 16
- 230000018044 dehydration Effects 0.000 claims abstract description 11
- 238000006297 dehydration reaction Methods 0.000 claims abstract description 11
- 230000008569 process Effects 0.000 claims abstract description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 39
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 26
- 239000000654 additive Substances 0.000 claims description 10
- 230000000996 additive effect Effects 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 7
- 238000004043 dyeing Methods 0.000 claims description 4
- 238000003745 diagnosis Methods 0.000 abstract description 7
- 210000003850 cellular structure Anatomy 0.000 abstract description 5
- 239000001993 wax Substances 0.000 description 21
- 235000019441 ethanol Nutrition 0.000 description 12
- 230000000694 effects Effects 0.000 description 8
- 239000004575 stone Substances 0.000 description 7
- 238000004519 manufacturing process Methods 0.000 description 4
- 210000004881 tumor cell Anatomy 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- 102000001554 Hemoglobins Human genes 0.000 description 2
- 108010054147 Hemoglobins Proteins 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000008034 disappearance Effects 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M sodium bicarbonate Substances [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 210000001015 abdomen Anatomy 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 230000006727 cell loss Effects 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 230000001744 histochemical effect Effects 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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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
-
- 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
- G01N1/286—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising
-
- 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
- G01N1/34—Purifying; Cleaning
-
- 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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- 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)
Abstract
The present invention relates to a kind of cell block based on courageous and upright Pleural effusions and preparation method thereof, it is related to biological technical field.The preparation method of the cell block based on courageous and upright Pleural effusions includes：Courageous and upright Pleural effusions are centrifuged into 4 5min for the first time under conditions of 1900 2100r/min, obtain the first precipitation.Erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process, and after lighter, second of centrifugation, the obtain second precipitation is wrapped in lens paper, fixed, dehydration waxdip, embedding.It is simple and convenient, make obtained cell block, cell component is more, and eucaryotic cell structure is complete, high power clear background, effectively improves the accuracy rate of diagnosis.
Description
Technical field
The present invention relates to biological technical field, and more particularly to a kind of cell block and its preparation based on courageous and upright Pleural effusions
Method.
Background technology
Exfoliative cytology inspection is one of diagnosing tumor effective means, a small amount of chest and abdomen drawn in common smear manufacturing process
Water cannot represent the situation of whole Pleural effusions, and make and finish remaining Pleural effusions often abandoned afterwards, and cell block can
To collect most cells in censorship Pleural effusions, to observe the form of most cells, and cell block is easy to preserve,
The Sensitivity and Specificity of cell pathology diagnosis is substantially increased, regular growth smear is compensate for and makes limited and not enough make more
The shortcoming of kind of dyeing, can further be studied and perspective study.
But the set time is grown during existing cell block makes, cell loss is obvious.Meanwhile in existing courageous and upright Pleural effusions
Due to there are substantial amounts of red blood cell, influencing the quality and diagnostic result of wax stone, some intractable cases needed to do cellular immunity
Also the judgement to positive findings can be influenced during histochemical stain.
The content of the invention
It is an object of the invention to provide a kind of cell block based on courageous and upright Pleural effusions, cell component is more, eucaryotic cell structure
Completely, high power clear background, effectively improves the accuracy rate of diagnosis.
It is simple and convenient another object of the present invention is to provide a kind of cell block based on courageous and upright Pleural effusions, have
Effect solves the above problems.
The present invention is solved its technical problem and is realized using following technical scheme.
The present invention proposes a kind of preparation method of the cell block based on courageous and upright Pleural effusions, it includes：
Courageous and upright Pleural effusions are centrifuged into 4-5min for the first time under conditions of 1900-2100r/min, obtain the first precipitation.
Erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process, and
After lighter, second of centrifugation, the obtain second precipitation is wrapped in lens paper, fixed, dehydration, waxdip, embedding.
The present invention proposes a kind of cell block based on courageous and upright Pleural effusions as made from above-mentioned preparation method.
The beneficial effect of the cell block based on courageous and upright Pleural effusions of the embodiment of the present invention and preparation method thereof is：
Courageous and upright Pleural effusions are centrifuged into 4-5min for the first time under conditions of 1900-2100r/min, effective enrichment of cell, is protected
Broken cell debris is removed while holding cell integrity.
Erythrocyte cracked liquid makes erythrocytolysis, the disappearance that interference diagnoses, and prevents the interference of red blood cell, effectively carries on the back high power
Scape centrifuges for second clearly simultaneously as after red blood cell is destroyed, and forms hemoglobin centrifugation more firm thin
Born of the same parents' stent, and by other tangible cells, especially tumour cell, enlist the services of makes it not be damaged in stent, makes last the
Two precipitations, which can be moved to as tissue block in lens paper, to be wrapped, it is not necessary to is added agar centrifugal treating again, is effectively simplified
Production process, while make the quality of wax stone more reliable.
After lighter, the red blood cell of second of centrifugation explanation overwhelming majority is destroyed, and carries out second of centrifugation behaviour at this time
Make, make the enrichment of remaining tangible cell progress again.Finally, it is fixed, is dehydrated, waxdip, embedding, makes obtained to be based on blood
Cell component is more in the cell block of property Pleural effusions, and eucaryotic cell structure is complete, high power clear background, effectively improves the accurate of diagnosis
Rate.
Embodiment
, below will be in the embodiment of the present invention to make the purpose, technical scheme and advantage of the embodiment of the present invention clearer
Technical solution be clearly and completely described.The person that is not specified actual conditions in embodiment, builds according to normal condition or manufacturer
The condition of view carries out.Reagents or instruments used without specified manufacturer, is the conventional production that can be obtained by commercially available purchase
Product.
Cell block based on courageous and upright Pleural effusions of the embodiment of the present invention and preparation method thereof is specifically described below.
The present invention provides a kind of cell block based on courageous and upright Pleural effusions, its cell component is more, and eucaryotic cell structure is complete, high power
Clear background, effectively improves the accuracy rate of diagnosis.It is made by following methods：
S1. courageous and upright Pleural effusions are centrifuged into 4-5min for the first time under conditions of 1900-2100r/min, obtains the first precipitation.
Since the cell quantity contained in courageous and upright Pleural effusions disperses, in order to make cell enrichment, make in obtained cell block
Cell component it is more, while improve producing efficiency, therefore, by courageous and upright Pleural effusions under conditions of 1900-2100r/min for the first time
4-5min is centrifuged, obtains the first precipitation.Complete cell is enriched with, keeps removing broken cell while cell integrity
Relic.
Preferably, before first time centrifuges, courageous and upright Pleural effusions are stood, make cell deposition in bottom, take bottom sample into
Row first time centrifugal treating, effectively improves the efficiency of centrifugation for the first time.
S2. erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process,
And after lighter, second of centrifugation, obtains the second precipitation.
Due to containing substantial amounts of red blood cell in courageous and upright Pleural effusions, and quality and diagnosis of the presence of red blood cell for wax stone
As a result, and while needing to do cell immunohistochemical staining to some intractable cases can also influence judgement to positive findings all
There are undesirable influence.Therefore, cracked using erythrocyte cracked liquid for the red blood cell in the first precipitation of enrichment so that
The high power clear background of the section of obtained wax stone, easy to accurately observe and diagnose.
It should be noted that gently being broken up when the first precipitation is mixed with erythrocyte cracked liquid, it is necessary to first be precipitated first, make
In first precipitation the red blood cell that is enriched be sufficiently mixed with erythrocyte cracked liquid uniformly, make its for red blood cell splitting action more preferably
Change.Preferably, by first precipitation gently break up using in adition process vibration make the first precipitation dissolving by the way of carry out, effectively
The efficiency of work is improved, while vibration makes contact of the cell with erythrocyte cracked liquid be more uniformly distributed and fully.
Alternatively, erythrocyte cracked liquid can be by NH4Cl, NaHCO3And EDTA configurations, wherein NH4Cl is split in red blood cell
The concentration solved in liquid is 0.15M, NaHCO3Concentration in erythrocyte cracked liquid is 0.01M, and EDTA is in erythrocyte splitting
Concentration in liquid is 1mM.
In preferred embodiments of the present invention, erythrocyte cracked liquid is the ethanol solution that volume fraction is 50%, wherein, volume
Fraction is 50% ethanol solution, and erythrocytolysis, the disappearance that interference can not only diagnosed, prevent the interference of red blood cell, have
Effect is while make high power clear background, after being destroyed due to red blood cell, centrifuges for second, make hemoglobin centrifugation formed compared with
For firm cytoskeleton, and by other tangible cells, especially tumour cell, enlist the services of makes it not be damaged in stent,
Make the second last precipitation to be moved to as tissue block in lens paper to be wrapped, it is not necessary to add agar and centrifuge place again
Reason, effectively simplifies production process, while makes the quality of wax stone more reliable.
Since red blood cell is destroyed, the mixed liquor in the first precipitation with erythrocyte cracked liquid is preferable in lighter
Afterwards, such as after being changed into pale red, illustrate that the red blood cell of the overwhelming majority is destroyed, carry out second of centrifugally operated at this time, make remaining
Tangible cell carries out enrichment again.
Alternatively, second centrifugation carries out 4-5min under conditions of 1900-2100r/min, and being effectively enriched with remaining has
Cell rupture is prevented while shape cell.
S3. the obtain second precipitation is wrapped in lens paper, fixed, dehydration, waxdip, embedding.
Wherein, the ethanol solution that the fixed fixer used is 90-95% for volume fraction, is preferably that volume fraction is
95% ethanol solution is, it is necessary to illustrate, ethanol solution refers both to ethanol water in the present invention.Wherein, the ethanol of 90-95%
Solution effectively separates out protein, suppresses cell and self-dissolving occurs, make its shape and internal structure, material composition etc. as far as possible with approximation
The state of live body preserves.
In preferred embodiments of the present invention, fixation includes：After second precipitation is mixed with acetone, in 1500-1650r/
Min, after centrifuging 4-5min, abandons supernatant, obtains the 3rd precipitation.
It should be noted that the second precipitation sample is uniformly mixed with acetone, it is necessary to first gently break up the second precipitation sample.
Preferably, ketone oil fills the tube wall of the second precipitation, is added along tube wall, makes fixed effect more abundant, fixed effect
Fruit is good, effectively prevents fixed uneven, causes the effect of cell block poor.
It is further preferred that the volume ratio of the additive amount of the second additive amount precipitated and acetone is 1:3-4.The additive amount
Under the conditions of, make the cell that the second precipitation is enriched with fully fixed by acetone.
It is further preferred that the temperature of acetone is 3.5-4.5 DEG C, effectively fixed cell while, prevent cellular contraction compared with
Greatly, cause cytomorphosis serious, and cyto-architectural imperfection, influence final effect.
Dehydration is included in 44-46 DEG C, is dehydrated 110-125min in the ethanol solution that volume fraction is 90-96%, then
It is dehydrated 1-2 times in absolute ethyl alcohol, is dehydrated 85-120min every time.It is fixed before dehydration, the 3rd precipitation is more readily formed
Cell block.
In the present invention, it is preferable that waxdip includes：By the 3rd wax of the precipitation coated on melt surface shape, toasted in 60-65 DEG C
4-6min.By toasting 4-6min in 60-65 DEG C, on the one hand further promote the effect of waxdip, wax is fully infiltrated on cell.
Specifically, by after the 3rd wax of the precipitation coated on melt surface shape, the 3rd precipitation is quickly flattened in the surface of wax,
20-25 DEG C is quickly cooled to, is toasted after standing 5-20min.
Using above method waxdip, cell block is located at the top of wax stone, on the one hand easily section, while cell knot under light microscopic
Structure is more complete, and cell and clear background, thickness is uniform, and mesothelium, tumour and inflammatory cell bright, cell are uniformly dispersed, cell
Clearly, caryoplasm staining versus is obvious, and cellular morphology is approximate with conventional organization making paraffin section HE, easily differentiates, easy to observe
Analysis, another aspect paraffin mass can preserve for a long time.
Waxdip can also be after 60-65 DEG C of first time waxdip 30-35min, after keeping the temperature 10-20min, then in 60-65
DEG C of second waxdip carries out 110-130min.Effectively improve the efficiency of waxdip.
Embedding uses the prior art, then this does not do and specifically repeats.Alternatively, after embedding, further include section with
And dyeing.Wherein, section for example can be that the wax stone of the courageous and upright Pleural effusions after embedding is switched to the thin slice that thickness is 3-6um, have
Effect solves the problems, such as that eucaryotic cell structure is not known caused by section is too thick.
The feature and performance of the present invention are described in further detail with reference to embodiments.
Embodiment 1
A kind of cell block based on courageous and upright Pleural effusions, it is made by following methods：
Courageous and upright Pleural effusions are centrifuged into 5min for the first time under conditions of 2000r/min, obtain the first precipitation.
Erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process, and
After lighter, second of centrifugation 5min under conditions of 2000r/min, the obtain second precipitation is wrapped in lens paper,
Fixed, dehydration waxdip, embedding, section and dyeing.
Wherein, erythrocyte cracked liquid is the ethanol solution that volume fraction is 50%.
Fixation includes：It is 1 to precipitate the acetone for being 4 DEG C with temperature according to additive amount volume ratio by second:After 3 mixing, in
1600r/min, after centrifuging 5min, abandons supernatant, obtains the 3rd precipitation.
Waxdip includes：After the 3rd wax of the precipitation coated on melt surface shape, the 3rd precipitation is quickly flattened in the table of wax
Face, is quickly cooled to 24 DEG C, stands 5min and toasts 5min after 65 DEG C.
Embodiment 2
A kind of cell block based on courageous and upright Pleural effusions, it is made by following methods：
Courageous and upright Pleural effusions are centrifuged into 5min for the first time under conditions of 2100r/min, obtain the first precipitation.
Erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process, and
After lighter, second of centrifugation 5min under conditions of 1900r/min, the obtain second precipitation is wrapped in lens paper,
Fixed, dehydration waxdip, embedding.
Wherein, erythrocyte cracked liquid is the ethanol solution that volume fraction is 50%.
Fixation includes：It is 1 to precipitate the acetone for being 3.5 DEG C with temperature according to additive amount volume ratio by second:After 3.5 mixing,
In 1650r/min, after centrifuging 4min, supernatant is abandoned, obtains the 3rd precipitation.
Waxdip includes：After the 3rd wax of the precipitation coated on melt surface shape, the 3rd precipitation is quickly flattened in the table of wax
Face, is quickly cooled to 25 DEG C, stands 15min and toasts 6min after 60 DEG C.
Embodiment 3
A kind of cell block based on courageous and upright Pleural effusions, it is made by following methods：
Courageous and upright Pleural effusions are centrifuged into 4.5min for the first time under conditions of 1900r/min, obtain the first precipitation.
Erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process, and
After lighter, second of centrifugation 4.5min under conditions of 2000r/min, lens paper is wrapped in by the obtain second precipitation
It is interior, fixed, dehydration waxdip, embedding.
Wherein, erythrocyte cracked liquid is the ethanol solution that volume fraction is 50%.
Fixation includes：It is 1 to precipitate the acetone for being 4.5 DEG C with temperature according to additive amount volume ratio by second:After 4 mixing, in
1500r/min, after centrifuging 4.5min, abandons supernatant, obtains the 3rd precipitation.
Waxdip includes：After the 3rd wax of the precipitation coated on melt surface shape, the 3rd precipitation is quickly flattened in the table of wax
Face, is quickly cooled to 23 DEG C, stands 10min and toasts 6min after 63 DEG C.
Embodiment 4
A kind of cell block based on courageous and upright Pleural effusions, it is made by following methods：
Courageous and upright Pleural effusions are centrifuged into 4.5min for the first time under conditions of 2000r/min, obtain the first precipitation.
Erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process, and
After lighter, second of centrifugation 4.5min under conditions of 2000r/min, lens paper is wrapped in by the obtain second precipitation
It is interior, fixed, dehydration waxdip, embedding.
Wherein, erythrocyte cracked liquid is the ethanol solution that volume fraction is 50%.
Fixation includes：It is 1 to precipitate the acetone for being 4 DEG C with temperature according to additive amount volume ratio by second:After 3.7 mixing, in
1600r/min, after centrifuging 5min, abandons supernatant, obtains the 3rd precipitation.
Waxdip includes：After the 3rd wax of the precipitation coated on melt surface shape, the 3rd precipitation is quickly flattened in the table of wax
Face, is quickly cooled to 23 DEG C, stands 17min and toasts 5min after 63 DEG C.
Embodiment 5
A kind of cell block based on courageous and upright Pleural effusions, it is made by following methods：
Courageous and upright Pleural effusions are centrifuged into 5min for the first time under conditions of 2000r/min, obtain the first precipitation.
Erythrocyte cracked liquid is slowly added in the first precipitation, and vibration dissolves the first precipitation in adition process, and
After lighter, second of centrifugation 4.5min under conditions of 2000r/min, lens paper is wrapped in by the obtain second precipitation
It is interior, fixed, dehydration waxdip, embedding.
Wherein, erythrocyte cracked liquid is the ethanol solution that volume fraction is 50%.
Fixation includes：It is 1 to precipitate the acetone for being 4 DEG C with temperature according to additive amount volume ratio by second:After 3.5 mixing, in
1650r/min, after centrifuging 5min, abandons supernatant, obtains the 3rd precipitation.
Waxdip includes：After the 3rd wax of the precipitation coated on melt surface shape, the 3rd precipitation is quickly flattened in the table of wax
Face, is quickly cooled to 23 DEG C, stands 13min and toasts 5min after 63 DEG C.
To sum up, the preparation method of the cell block provided in an embodiment of the present invention based on courageous and upright Pleural effusions, it is easy to operate just
Victory, the set time is shorter, and the almost all of cell in effective fixed sample, prevents the waste of cell, while effectively destroys red
Cell, makes that obtained cell block cell component is more, and eucaryotic cell structure is complete, and high power clear background, effectively improves the accurate of diagnosis
Rate.
Embodiments described above is part of the embodiment of the present invention, instead of all the embodiments.The reality of the present invention
The detailed description for applying example is not intended to limit the scope of claimed invention, but is merely representative of the selected implementation of the present invention
Example.Based on the embodiments of the present invention, those of ordinary skill in the art are obtained without creative efforts
Every other embodiment, belongs to the scope of protection of the invention.
Claims (10)
- A kind of 1. preparation method of the cell block based on courageous and upright Pleural effusions, it is characterised in that including：The courageous and upright Pleural effusions are centrifuged into 4-5min for the first time under conditions of 1900-2100r/min, obtain the first precipitation；Erythrocyte cracked liquid is slowly added in the described first precipitation, and vibration makes first precipitation molten in adition process Solution, and after lighter, second of centrifugation, the obtain second precipitation is wrapped in lens paper, fixed, dehydration, waxdip, bag Bury.
- 2. preparation method according to claim 1, it is characterised in that the erythrocyte cracked liquid is that volume fraction is 50% Ethanol solution.
- 3. preparation method according to claim 1, it is characterised in that fixation includes：Described second precipitation is mixed with acetone After conjunction, in 1500-1650r/min, after centrifuging 4-5min, supernatant is abandoned, obtains the 3rd precipitation.
- 4. preparation method according to claim 3, it is characterised in that the additive amount and the acetone of second precipitation The volume ratio of additive amount is 1:3-4.
- 5. preparation method according to claim 3, it is characterised in that the temperature of the acetone is 3.5-4.5 DEG C.
- 6. preparation method according to claim 3, it is characterised in that waxdip includes：Described 3rd precipitation is coated on table The wax of face molten, 4-6min is toasted in 60-65 DEG C.
- 7. preparation method according to claim 6, it is characterised in that by the described 3rd precipitation coated on melt surface shape After wax, by the quick pressing of the described 3rd precipitation in the surface of the wax, 20-25 DEG C is quickly cooled to, is dried after standing 5-20min It is roasting.
- 8. preparation method according to claim 1, it is characterised in that second of centrifugation is in the condition of 1900-2100r/min Lower carry out 4-5min.
- 9. preparation method according to claim 1, it is characterised in that after embedding, further include section and dyeing.
- 10. the cell block based on courageous and upright Pleural effusions made from the preparation method as described in claim 1-9 any one.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109975090A (en) * | 2019-04-15 | 2019-07-05 | 马晓丽 | A kind of preparation method of thyroid gland, mammary gland Fine-needle puncture cell tissue block |
| CN112082836A (en) * | 2020-09-15 | 2020-12-15 | 海南医学院第一附属医院 | Clinical bloody pleural effusion cell wax block and preparation method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112082836A (en) * | 2020-09-15 | 2020-12-15 | 海南医学院第一附属医院 | Clinical bloody pleural effusion cell wax block and preparation method thereof |
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