CN108535077A - A kind of pap staining liquid and application process - Google Patents
A kind of pap staining liquid and application process Download PDFInfo
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- CN108535077A CN108535077A CN201810314097.1A CN201810314097A CN108535077A CN 108535077 A CN108535077 A CN 108535077A CN 201810314097 A CN201810314097 A CN 201810314097A CN 108535077 A CN108535077 A CN 108535077A
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- dyeing
- liquid
- pap staining
- alcohol
- staining liquid
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- 239000007788 liquid Substances 0.000 title claims abstract description 33
- 238000010186 staining Methods 0.000 title claims abstract description 17
- 238000000034 method Methods 0.000 title claims description 14
- 238000004043 dyeing Methods 0.000 claims abstract description 46
- 230000008685 targeting Effects 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 47
- 235000019441 ethanol Nutrition 0.000 claims description 22
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 18
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 16
- 238000004140 cleaning Methods 0.000 claims description 12
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 claims description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 229960000583 acetic acid Drugs 0.000 claims description 8
- 239000012362 glacial acetic acid Substances 0.000 claims description 8
- 239000008213 purified water Substances 0.000 claims description 8
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 6
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 6
- 239000007853 buffer solution Substances 0.000 claims description 5
- 210000000805 cytoplasm Anatomy 0.000 claims description 5
- 229960001506 brilliant green Drugs 0.000 claims description 4
- HXCILVUBKWANLN-UHFFFAOYSA-N brilliant green cation Chemical compound C1=CC(N(CC)CC)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](CC)CC)C=C1 HXCILVUBKWANLN-UHFFFAOYSA-N 0.000 claims description 4
- NALMPLUMOWIVJC-UHFFFAOYSA-N n,n,4-trimethylbenzeneamine oxide Chemical compound CC1=CC=C([N+](C)(C)[O-])C=C1 NALMPLUMOWIVJC-UHFFFAOYSA-N 0.000 claims description 4
- 229940032753 sodium iodate Drugs 0.000 claims description 4
- 235000015281 sodium iodate Nutrition 0.000 claims description 4
- 239000011697 sodium iodate Substances 0.000 claims description 4
- 239000000243 solution Substances 0.000 claims description 4
- PQMOXTJVIYEOQL-UHFFFAOYSA-N Cumarin Natural products CC(C)=CCC1=C(O)C(C(=O)C(C)CC)=C(O)C2=C1OC(=O)C=C2CCC PQMOXTJVIYEOQL-UHFFFAOYSA-N 0.000 claims description 2
- FSOGIJPGPZWNGO-UHFFFAOYSA-N Meomammein Natural products CCC(C)C(=O)C1=C(O)C(CC=C(C)C)=C(O)C2=C1OC(=O)C=C2CCC FSOGIJPGPZWNGO-UHFFFAOYSA-N 0.000 claims description 2
- 239000007983 Tris buffer Substances 0.000 claims description 2
- 229940037003 alum Drugs 0.000 claims description 2
- ZYGHJZDHTFUPRJ-UHFFFAOYSA-N coumarin Chemical compound C1=CC=C2OC(=O)C=CC2=C1 ZYGHJZDHTFUPRJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000008055 phosphate buffer solution Substances 0.000 claims description 2
- IYDGMDWEHDFVQI-UHFFFAOYSA-N phosphoric acid;trioxotungsten Chemical compound O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.O=[W](=O)=O.OP(O)(O)=O IYDGMDWEHDFVQI-UHFFFAOYSA-N 0.000 claims description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical group OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 abstract description 2
- 238000004040 coloring Methods 0.000 description 12
- 239000000975 dye Substances 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 238000010827 pathological analysis Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 3
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 2
- ZXRRHFSTAFVGOC-UHFFFAOYSA-N [AlH3].[K] Chemical compound [AlH3].[K] ZXRRHFSTAFVGOC-UHFFFAOYSA-N 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 239000001048 orange dye Substances 0.000 description 2
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- UYDPQDSKEDUNKV-UHFFFAOYSA-N phosphanylidynetungsten Chemical compound [W]#P UYDPQDSKEDUNKV-UHFFFAOYSA-N 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000012128 staining reagent Substances 0.000 description 1
- 238000011121 vaginal smear Methods 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
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of pap staining liquid and its applications, belong to biotechnology.Specifically the pap staining liquid includes nuclear targeting liquid and endochylema dyeing liquor.Pap staining liquid provided by the invention is easy to operate, even dyeing, and the eucaryotic cell structure after dyeing is clear, and cell color is bright-coloured, is easy to Clinicopathologic Diagnosis observation and judges.
Description
Technical field
The present invention relates to biotechnology, more particularly to a kind of pap staining liquid, while the invention further relates to the dyeing
Liquid application process.
Background technology
During current medical diagnosis on disease, pathological diagnosis is the important sources of diagnostic message, has important clinical significance.
It is also most common colouring method that Papanicolaou's vaginal smear technique, which is best in pathological section and cast-off cells dyeing, and the colouring method is not only
It is clear with real nuclear structures, the features such as color separation is apparent, and transparency is good, and endochylema is bright-coloured by color, moreover it is possible to make sample after processing
It is not easy to decolourize, can preserve for a long time.Pap staining reagent includes haematoxylin dye liquor, orange dye liquor, EA50 dye liquors and 95% ethyl alcohol,
100% ethyl alcohol, absolute ethyl alcohol etc.；Papanicolaou stain procedure flow is：95% ethyl alcohol is fixed（15min）→ clear water rinses repeatedly → bush
Uniformly dyeing liquid（3-5min）→ clear water flushes three times → the ethyl alcohol rinsing of oil blackeite → 95% → orange dye liquor（1-10s）→ 95% ethyl alcohol rinses
→ 95% ethyl alcohol rinses → 100% ethyl alcohol rinsing → EA50（3-5m）→ 95% ethyl alcohol rinses → 95% ethyl alcohol rinsing → absolute ethyl alcohol drift
Wash → absolute ethyl alcohol（2min）→ dimethylbenzene（2min）→ neutral gum mounting.
Traditional Pasteur's color liquid configuration is complicated, and dyeing formality is complicated, and dyeing time is longer, when coloration result is dyed
Between, rinsing time, many factors such as rinsing times influence, and effect is unstable, influences the judgement of pathological diagnosis.
Invention content
The present invention is to solve above-mentioned deficiency of the prior art, provides a kind of pap staining liquid of rapid dyeing and answers
Use method.Pap staining liquid configuration is easy, and staining procedure is brief, and dye after stain is stablized, easy to operate, color separation and coloring
Significantly.
Specifically：The pap staining liquid contains nuclear targeting liquid and cytoplasm dyeing liquor.
Nuclear targeting liquid is improvement haematoxylin dyeing liquid, and every liter of dyeing liquor its group is divided into：50 ~ 150ml of alcohol, glacial acetic acid 5 ~
40ml, 1 ~ 4g of hematoxylin, 2 ~ 25g of alum, 0.05 ~ 0.5g of sodium iodate, surplus are purified water.
Cytoplasm dyeing liquor is OG-EA dyeing liquors, and every liter of dye liquor its group is divided into：0.1 ~ 1g of brilliant green, the micro- yellow 2 ~ 6g of eosin, nothing
400 ~ 700ml of water-ethanol, 0.5 ~ 4g of phosphotungstic acid, 100 ~ 450ml of alcohol, 5 ~ 40ml of glacial acetic acid, buffer solution 2 ~ 50ml, orange 2 ~ 8g,
Surplus is purified water.
Alcohol in above-mentioned dyeing liquor is：One or more of methanol, ethyl alcohol, propyl alcohol, isopropanol and cumarin combine；On
It is Tris buffer solutions, phosphate buffer solution, one or more of citric acid solution group to state the buffer solution in dyeing liquor
It closes.
The application method of the pap staining liquid is：
1）After the completion of cytology film-making, wet fixation is waited for；
2）After flowing water cleaning nuclear targeting is carried out using the improvement haematoxylin dyeing liquid 1 ~ 2 minute；
3）Endochylema is carried out after flowing water cleaning using the OG-EA dyeing liquors to dye 2 ~ 3 minutes；
4）Graded ethanol cleaning and dewatering dries rear mounting；
5）Diagosis sentence read result under mirror.
Pap staining liquid provided by the invention, it is fast to nucleus and cytoplasm dyeing coloring respectively, it can complete within 1 ~ 2 minute
Coloring very well rinses loose colour and facilitates succinct, and staining procedure is brief, and dye after stain is stablized, easy to operate, and color separation is aobvious with coloring
It writes.
Description of the drawings
1 embodiment of attached drawing, one microscopy picture.
2 embodiment of attached drawing, two microscopy picture.
Specific implementation mode
Embodiment one：
Improve haematoxylin dyeing liquid 1, every liter of dyeing liquor its group is divided into：Methanol 100ml, glacial acetic acid 20ml, hematoxylin 2g, sulfuric acid
Aluminium potassium 10g, sodium iodate 0.1g, purified water 870ml.
OG-EA dyeing liquors 1, every liter of dye liquor its group are divided into：Brilliant green 0.5g, eosin micro- Huang 2.5g, absolute ethyl alcohol 500ml, phosphorus
Wolframic acid 1g, ethyl alcohol 200ml, glacial acetic acid 20ml, phosphate buffer 12ml, orange 3g, purified water 265ml.
Colouring method：
1）After the completion of cytology film-making, wet fixation is waited for；
2）After flowing water cleaning nuclear targeting is carried out using above-mentioned improvement haematoxylin dyeing liquid 1 1.5 minutes；
3）Endochylema is carried out after flowing water cleaning using above-mentioned OG-EA dyeing liquors 1 to dye 2.5 minutes；
4）Graded ethanol cleaning and dewatering dries rear mounting；
5）Diagosis sentence read result under mirror.
Entire dyeing course is completed for 8 minutes.Observe the effect of dyeing under the microscope after the completion：Slide cell compartment background
Clean free from admixture, cell dyeing is uniform, and coloring is bright-coloured, and color separation is apparent with coloring, is easy to pathological diagnosis judgement.
Embodiment two：
Improve haematoxylin dyeing liquid 2, every liter of dyeing liquor its group is divided into：Propyl alcohol 120ml, glacial acetic acid 30ml, hematoxylin 3g, sulfuric acid
Aluminium potassium 15g, sodium iodate 0.2g, purified water 835ml.
OG-EA dyeing liquors 2, every liter of dye liquor its group are divided into：Brilliant green 0.6g, eosin micro- Huang 3g, absolute ethyl alcohol 600ml, phosphorus tungsten
Sour 2g, isopropanol 300ml, glacial acetic acid 25ml, Tirs buffer solution 20ml, orange 4g, purified water 46ml.
Colouring method：
1）After the completion of cytology film-making, wet fixation is waited for；
2）After flowing water cleaning nuclear targeting is carried out using above-mentioned improvement haematoxylin dyeing liquid 21 minute；
3）Endochylema is carried out after flowing water cleaning using above-mentioned OG-EA dyeing liquors 2 to dye 2 minutes；
4）Graded ethanol cleaning and dewatering dries rear mounting；
5）Diagosis sentence read result under mirror.
Entire dyeing course is completed for 7 minutes.Observe the effect of dyeing under the microscope after the completion：Slide cell compartment background
Clean free from admixture, cell dyeing is uniform, and coloring is bright-coloured, and color separation is apparent with coloring, is easy to pathological diagnosis judgement.
Claims (8)
1. a kind of pap staining liquid, it is characterised in that：Contain nuclear targeting liquid and cytoplasm dyeing liquor.
2. pap staining liquid according to claim 1, it is characterised in that nuclear targeting liquid is improvement haematoxylin dyeing liquid.
3. pap staining liquid according to claim 1, it is characterised in that cytoplasm dyeing liquor is OG-EA dyeing liquors.
4. improvement haematoxylin dyeing liquid according to claim 2, it is characterised in that every liter of dye liquor its group is divided into：Alcohol 50 ~
150ml, 5 ~ 40ml of glacial acetic acid, 1 ~ 4g of hematoxylin, 2 ~ 25g of alum, 0.05 ~ 0.5g of sodium iodate, surplus are purified water.
5. OG-EA dyeing liquors according to claim 3, it is characterised in that every liter of dye liquor its group is divided into：0.1 ~ 1g of brilliant green, daybreak
Red micro- yellow 2 ~ 6g, 400 ~ 700ml of absolute ethyl alcohol, 0.5 ~ 4g of phosphotungstic acid, 100 ~ 450ml of alcohol, 5 ~ 40ml of glacial acetic acid, buffer solution 2 ~
50ml, orange 2 ~ 8g, surplus are purified water.
6. the OG-EA dyeing liquors described in improvement haematoxylin dyeing liquid according to claim 4 and claim 5, feature
It is that the alcohol in component is the combination of one or more of methanol, ethyl alcohol, propyl alcohol, isopropanol and cumarin.
7. OG-EA dyeing liquors according to claim 5, it is characterised in that the buffer solution in component is Tris buffer solutions,
Phosphate buffer solution, the combination of one or more of citric acid solution.
8. its application process of pap staining liquid according to claim 1 is：
1）After the completion of cytology film-making, wet fixation is waited for；
2）After flowing water cleaning nuclear targeting is carried out using the improvement haematoxylin dyeing liquid 1 ~ 2 minute；
3）Endochylema is carried out after flowing water cleaning using the OG-EA dyeing liquors to dye 2 ~ 3 minutes；
4）Graded ethanol cleaning and dewatering dries rear mounting；
5）Diagosis sentence read result under mirror.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810314097.1A CN108535077A (en) | 2018-04-10 | 2018-04-10 | A kind of pap staining liquid and application process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810314097.1A CN108535077A (en) | 2018-04-10 | 2018-04-10 | A kind of pap staining liquid and application process |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108535077A true CN108535077A (en) | 2018-09-14 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810314097.1A Pending CN108535077A (en) | 2018-04-10 | 2018-04-10 | A kind of pap staining liquid and application process |
Country Status (1)
| Country | Link |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110006724A (en) * | 2019-04-17 | 2019-07-12 | 郑州安图生物工程股份有限公司 | Trichomonad reagent is detected using Pasteur and Gram-staining process |
| CN112781963A (en) * | 2020-12-30 | 2021-05-11 | 深路医学科技(武汉)有限公司 | Papanicolaou staining solution and preparation method and staining method thereof |
| CN112798385A (en) * | 2020-12-30 | 2021-05-14 | 深圳市鹏一医疗仪器有限公司 | Environment-friendly hematoxylin staining solution, papanicolaou staining solution, and preparation methods and applications thereof |
-
2018
- 2018-04-10 CN CN201810314097.1A patent/CN108535077A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110006724A (en) * | 2019-04-17 | 2019-07-12 | 郑州安图生物工程股份有限公司 | Trichomonad reagent is detected using Pasteur and Gram-staining process |
| CN110006724B (en) * | 2019-04-17 | 2021-06-04 | 郑州安图生物工程股份有限公司 | Reagent for detecting trichomonas by adopting papanicolaou and gram staining method |
| CN112781963A (en) * | 2020-12-30 | 2021-05-11 | 深路医学科技(武汉)有限公司 | Papanicolaou staining solution and preparation method and staining method thereof |
| CN112798385A (en) * | 2020-12-30 | 2021-05-14 | 深圳市鹏一医疗仪器有限公司 | Environment-friendly hematoxylin staining solution, papanicolaou staining solution, and preparation methods and applications thereof |
| CN112781963B (en) * | 2020-12-30 | 2024-04-30 | 深路医学科技(武汉)有限公司 | Papanicolaou staining solution and preparation method and staining method thereof |
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Application publication date: 20180914 |