CN107490506B - The immediate processing method of pathological diagnosis kit and tissue - Google Patents
The immediate processing method of pathological diagnosis kit and tissue Download PDFInfo
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- CN107490506B CN107490506B CN201710651672.2A CN201710651672A CN107490506B CN 107490506 B CN107490506 B CN 107490506B CN 201710651672 A CN201710651672 A CN 201710651672A CN 107490506 B CN107490506 B CN 107490506B
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- 238000010827 pathological analysis Methods 0.000 title claims abstract description 45
- 238000003672 processing method Methods 0.000 title claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 96
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 claims abstract description 88
- 229960000935 dehydrated alcohol Drugs 0.000 claims abstract description 46
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 43
- 239000012153 distilled water Substances 0.000 claims abstract description 35
- 230000007935 neutral effect Effects 0.000 claims abstract description 26
- 238000012545 processing Methods 0.000 claims abstract description 22
- 239000004568 cement Substances 0.000 claims abstract description 18
- UKZQEOHHLOYJLY-UHFFFAOYSA-M ethyl eosin Chemical compound [K+].CCOC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C([O-])=C(Br)C=C21 UKZQEOHHLOYJLY-UHFFFAOYSA-M 0.000 claims abstract description 17
- 238000005406 washing Methods 0.000 claims abstract description 17
- XGZVUEUWXADBQD-UHFFFAOYSA-L lithium carbonate Chemical compound [Li+].[Li+].[O-]C([O-])=O XGZVUEUWXADBQD-UHFFFAOYSA-L 0.000 claims abstract description 16
- 229910052808 lithium carbonate Inorganic materials 0.000 claims abstract description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 51
- 239000011521 glass Substances 0.000 claims description 28
- 238000011010 flushing procedure Methods 0.000 claims description 27
- 229960000583 acetic acid Drugs 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 25
- 239000012362 glacial acetic acid Substances 0.000 claims description 24
- 238000003756 stirring Methods 0.000 claims description 15
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 claims description 14
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 claims description 14
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical compound CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 claims description 14
- BKIMMITUMNQMOS-UHFFFAOYSA-N nonane Chemical compound CCCCCCCCC BKIMMITUMNQMOS-UHFFFAOYSA-N 0.000 claims description 14
- 229910000474 mercury oxide Inorganic materials 0.000 claims description 13
- UKWHYYKOEPRTIC-UHFFFAOYSA-N mercury(ii) oxide Chemical compound [Hg]=O UKWHYYKOEPRTIC-UHFFFAOYSA-N 0.000 claims description 13
- 238000002360 preparation method Methods 0.000 claims description 13
- 238000010438 heat treatment Methods 0.000 claims description 12
- 206010016825 Flushing Diseases 0.000 claims description 11
- AMQJEAYHLZJPGS-UHFFFAOYSA-N N-Pentanol Chemical compound CCCCCO AMQJEAYHLZJPGS-UHFFFAOYSA-N 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 7
- 239000005457 ice water Substances 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- TVMXDCGIABBOFY-UHFFFAOYSA-N octane Chemical compound CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 claims description 7
- 239000003921 oil Substances 0.000 claims description 7
- 230000003902 lesion Effects 0.000 claims description 6
- 238000007790 scraping Methods 0.000 claims description 6
- 206010006223 Breast discharge Diseases 0.000 claims description 3
- 206010036790 Productive cough Diseases 0.000 claims description 3
- 210000004908 prostatic fluid Anatomy 0.000 claims description 3
- 230000028327 secretion Effects 0.000 claims description 3
- 210000003802 sputum Anatomy 0.000 claims description 3
- 208000024794 sputum Diseases 0.000 claims description 3
- 210000002700 urine Anatomy 0.000 claims description 3
- 235000004443 Ricinus communis Nutrition 0.000 claims 1
- 240000000528 Ricinus communis Species 0.000 claims 1
- 238000009835 boiling Methods 0.000 claims 1
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 17
- 238000000034 method Methods 0.000 abstract description 16
- 230000008569 process Effects 0.000 abstract description 9
- 230000007170 pathology Effects 0.000 abstract description 4
- 238000011835 investigation Methods 0.000 abstract description 3
- 210000001519 tissue Anatomy 0.000 description 59
- 239000000243 solution Substances 0.000 description 40
- 239000000975 dye Substances 0.000 description 37
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 23
- 235000019441 ethanol Nutrition 0.000 description 23
- 229960004756 ethanol Drugs 0.000 description 21
- 230000000694 effects Effects 0.000 description 10
- 206010028980 Neoplasm Diseases 0.000 description 8
- 102000004169 proteins and genes Human genes 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 238000001035 drying Methods 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- 239000004359 castor oil Substances 0.000 description 6
- 235000019438 castor oil Nutrition 0.000 description 6
- 239000003292 glue Substances 0.000 description 6
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 6
- 230000003321 amplification Effects 0.000 description 5
- 210000000805 cytoplasm Anatomy 0.000 description 5
- 230000018044 dehydration Effects 0.000 description 5
- 238000006297 dehydration reaction Methods 0.000 description 5
- 238000003199 nucleic acid amplification method Methods 0.000 description 5
- 239000001044 red dye Substances 0.000 description 5
- 238000002604 ultrasonography Methods 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- 238000003912 environmental pollution Methods 0.000 description 4
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 4
- 229910052737 gold Inorganic materials 0.000 description 4
- 239000010931 gold Substances 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 4
- 238000009595 pap smear Methods 0.000 description 4
- 230000001575 pathological effect Effects 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 241001062009 Indigofera Species 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000383 hazardous chemical Substances 0.000 description 3
- 239000004615 ingredient Substances 0.000 description 3
- 210000001685 thyroid gland Anatomy 0.000 description 3
- 239000008096 xylene Substances 0.000 description 3
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- 208000032843 Hemorrhage Diseases 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 206010048612 Hydrothorax Diseases 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- 239000000908 ammonium hydroxide Substances 0.000 description 2
- 150000001450 anions Chemical class 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 125000001309 chloro group Chemical group Cl* 0.000 description 2
- 239000003086 colorant Substances 0.000 description 2
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 2
- 238000012864 cross contamination Methods 0.000 description 2
- 230000001086 cytosolic effect Effects 0.000 description 2
- 230000004069 differentiation Effects 0.000 description 2
- 239000004519 grease Substances 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 238000009940 knitting Methods 0.000 description 2
- 210000001165 lymph node Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 125000000896 monocarboxylic acid group Chemical group 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 210000002445 nipple Anatomy 0.000 description 2
- 231100000989 no adverse effect Toxicity 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 239000001509 sodium citrate Substances 0.000 description 2
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 2
- 238000005728 strengthening Methods 0.000 description 2
- QBUKAFSEUHGMMX-MTJSOVHGSA-N (5z)-5-[[3-(1-hydroxyethyl)thiophen-2-yl]methylidene]-10-methoxy-2,2,4-trimethyl-1h-chromeno[3,4-f]quinolin-9-ol Chemical group C1=CC=2NC(C)(C)C=C(C)C=2C2=C1C=1C(OC)=C(O)C=CC=1O\C2=C/C=1SC=CC=1C(C)O QBUKAFSEUHGMMX-MTJSOVHGSA-N 0.000 description 1
- CMCZKWJUYJOKFX-IWIPYMOSSA-N (5z)-5-[[3-(1-hydroxypentyl)thiophen-2-yl]methylidene]-10-methoxy-2,2,4-trimethyl-1h-chromeno[3,4-f]quinolin-9-ol Chemical group C1=CSC(\C=C/2C3=C4C(C)=CC(C)(C)NC4=CC=C3C3=C(OC)C(O)=CC=C3O\2)=C1C(O)CCCC CMCZKWJUYJOKFX-IWIPYMOSSA-N 0.000 description 1
- 240000008067 Cucumis sativus Species 0.000 description 1
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 description 1
- WHXSMMKQMYFTQS-UHFFFAOYSA-N Lithium Chemical compound [Li] WHXSMMKQMYFTQS-UHFFFAOYSA-N 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- SEACYXSIPDVVMV-UHFFFAOYSA-L eosin Y Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C([O-])=C(Br)C=C21 SEACYXSIPDVVMV-UHFFFAOYSA-L 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 230000036571 hydration Effects 0.000 description 1
- 238000006703 hydration reaction Methods 0.000 description 1
- 229960002163 hydrogen peroxide Drugs 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229910052744 lithium Inorganic materials 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 238000012805 post-processing Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 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
- G01N1/34—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
-
- 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
- G01N1/42—Low-temperature sample treatment, e.g. cryofixation
-
- 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
-
- 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
- G01N2001/305—Fixative compositions
- G01N2001/307—Fixative compositions non-toxic, no Hg, no formaldehyde
-
- 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
- G01N2001/364—Embedding or analogous mounting of samples using resins, epoxy
Abstract
The present invention relates to the kit of pathological diagnosis and its application method, which includes four washing bottles with identification label, wherein headpin is equipped with Harri hematoxylin dye liquor and ClarkeShi fixer；No. 2 bottled spirit soluble eosin Y, dehydrated alcohol, distilled water and lithium carbonate；No. 3 bottled clarifier；No. 4 bottled neutral Instant cements having for moist mounting.The time needed for existing tissue is handled can be greatly reduced using the kit, reduce the process links of processing and reduces reagent dosage, conducive to the development to medical services from far-off regions, it can accomplish to check on the spot in volunteer medical consultation from far-off regions, generaI investigation, there is pathology diagnostic result, on the spot conducive to the raising of medical service level from far-off regions.
Description
Technical field
The present invention relates to the kit of pathological diagnosis and its application methods, fixation, dehydration and dyeing including tissue.
Background technique
In the prior art, a kind of method for having the kit for pathologic diagnosis of tumor and histotomy being dyed,
Its application number: 201310152840.5, the applying date: 2013-04-27, publication number: CN103245789A, publication date: 2013-08-
14, this is used for the kit of pathologic diagnosis of tumor, including the probe for pathologic diagnosis of tumor；The probe includes gold nano
The antibody and Coomassie brilliant blue of cluster, tumour-specific mark, the gold nanoclusters and the tumour-specific mark resist
Body is connected by click chemistry mode, the Coomassie brilliant blue and the antibody for being connected with the tumour-specific mark
Gold nanoclusters are combined by electrostatic means；Wherein, the gold nanoclusters, the tumour-specific mark antibody and described examine
The molar ratio of Mas bright blue is 1~10:1~100:1~100.
The method dyed using the kit to histotomy, is included the following steps:
1) by frozen tissue section or in advance, the paraffin tissue sections after the hydration process that dewaxes successively are soaked in anhydrous second
In the ethanol solution that the ethanol solution and mass fraction that alcohol, mass fraction are 95% are 70%, 5 minutes every time, then use
PBS washing；
2) hydrogenperoxide steam generator that volume fraction is 3% is added dropwise on tissue sections, is stored at room temperature 10 minutes, and through PBS
Washing；
3) histotomy is put into 95 DEG C of sodium citrate buffer solutions constant temperature to impregnate 10~15 minutes, and through PBS
Washing, wherein the concentration of sodium citrate buffer solution is 0.01mol/L；
4) sealer is added dropwise on tissue sections, after closing 20 minutes at room temperature, gets rid of surplus liquid；
5) probe for being used for pathologic diagnosis of tumor is added dropwise on tissue sections, stands 1 hour, 4 DEG C and stays overnight at room temperature
Or 1 hour is stood at 37 DEG C, and wash through PBS, wherein the final concentration of 0.01-10mmol/L of the probe；
6) nuclear stain is added dropwise on tissue sections, is then incubated for 7-10 minutes at 37 DEG C, makes the final concentration of of nuclear stain
0.1-10 μ g/ml, and through PBS or originally water washing；Mounting, then microscopy.
Health ministry entrusts " the clinical technique specification pathology organized the compilation by pathology branch, Chinese Medical Association
Credit volume " describe the existing process flow of the applicable tissue of the present invention:
(i) the processing of smear:
1. being directly soaked in fixer after slightly drying, at least 15-30min；
2. dimethylbenzene I: 5-10min；
3. dimethylbenzene II: 5-10min；(purpose through two step xylene soaks is: being more conducive to will be remaining in tissue
Previous step reagent and foreign matter are removed)
4. dehydrated alcohol I: 1-3min；
5. dehydrated alcohol II: 1-3min；(purpose through two step soaked in absolute ethyl alcohol is: being more conducive to will be in tissue
Moisture is taken away)
6. 95% ethyl alcohol: 1-3min；
7. 95% ethyl alcohol: 1-3min；(purpose impregnated through two steps, 95% ethyl alcohol is: strengthening 95% ethyl alcohol in tissue
In penetration power, taken away conducive to by the moisture in tissue)
8. 80% ethyl alcohol: 1min；
9. distilled water: 1min；
10. bush sperm dyes: 5-10min；
11. flowing water washes away bush sperm: 1min；
12. 1% hydrochloric acid of volumetric concentration-ethyl alcohol: 1-3s；
13. soft washing: 1-2s；
14. returning blue (with warm water or 1% ammonium hydroxide etc.): 5-10s；
15. flowing water rinses: 1-2min；
16. distilled water is slightly washed: 1-2min；
17.0.5% Yihong liquid dyes: 1-3min；
18. distilled water is slightly washed: 1-2s；
19.80% ethyl alcohol: 1-2s；
20. 95% ethyl alcohol I: 2-3min；
21. 95% ethyl alcohol II: 2-3min；(purpose impregnated through two steps, 95% ethyl alcohol is: strengthening 95% ethyl alcohol and is organizing
Penetration power in piece is taken away conducive to by the moisture in tissue)
22. dehydrated alcohol I: 3-5min；
23. dehydrated alcohol II: 3-5min；(purpose through two step soaked in absolute ethyl alcohol is: being more conducive to will be in tissue
Moisture is taken away)
24. carbolic acid-dimethylbenzene: 3-5min；
25. dimethylbenzene I: 3-5min；
26. dimethylbenzene II: 3-5min；(purpose through two step xylene soaks is: being more conducive to will be remaining in tissue
Previous step reagent and foreign matter are removed)
27. neutral gum sealing.
Above-mentioned 13 and 14 can save, but 15 flushing periods need to extend to 10-15min；24 Xiang Keyong dehydrated alcohol generations
It replaces, northern area can omit.
(ii) frozen section is handled:
1. frozen section is fixed: 10-30s；
2. soft washing: 1-2s；
3. bush sperm dyes (60 DEG C): 30-60s；
4. flowing water washes away bush sperm: 5-10s；
5. 1% hydrochloric acid of volumetric concentration-ethyl alcohol: 1-3s；
6. soft washing: 1-2min；
7. returning blue (with warm water or 1% ammonium hydroxide etc.): 5-10s；
8. flowing water rinses: 15-30s；
9.0.5% Yihong liquid dyes: 1-2min；
10. distilled water is slightly washed: 1-2min；
11.80% ethyl alcohol: 1-2min；
12.95% ethyl alcohol: 1-2min；
13. dehydrated alcohol I: 1-2min；
14. dehydrated alcohol II: 1-2min；(purpose through two step soaked in absolute ethyl alcohol is: being more conducive to will be in tissue
Moisture is taken away)
15. carbolic acid-dimethylbenzene: 2-3min；
16. dimethylbenzene I: 2-3min；
17. dimethylbenzene II: 2-3min；(purpose through two step xylene soaks is: being more conducive to will be remaining in tissue
Previous step reagent and foreign matter are removed)
18. neutral gum sealing.
Above-mentioned 7 and 8 can save, but 9 flushing periods need to extend to 10-15min；15 Xiang Keyong dehydrated alcohols replace,
Northern area can omit.
It is in place of the deficiencies in the prior art: the conventional treatment of tissue used in pathological diagnosis but process tedious at present, step
It is rapid various；Required film-making container is more, occupied space；There are also cross contamination when in special circumstances, influence pathological diagnosis as a result, this
Outside, the Cucumber in reagent (such as dimethylbenzene) has certain toxicity, can cause environmental pollution and influence operator's body and mind
Health, these all constrain the extensive development of pathological diagnosis.
Summary of the invention
The object of the present invention is to provide a kind of pathological diagnosis kit and the immediate processing methods of tissue, existing to solve
Organized piece fixes, be dehydrated and time-consuming in dyeing course, and consumption reagent is more, takes up space greatly and the problem of environmental pollution；
It is convenient for carrying with occupying little space, pathological diagnosis tissue short processing time and can be used for different type pathological diagnosis group
The characteristics of knitting piece processing.
The object of the present invention is achieved like this: a kind of pathological diagnosis pathological diagnosis kit is provided with insighted
Four washing bottles not marked, wherein
Headpin: Harri hematoxylin dye liquor 85-95ml and ClarkeShi fixer 38-42ml is housed；
No. 2 bottles: spirit soluble eosin Y 0.38-0.42g, dehydrated alcohol 105-115ml, distilled water 9-12ml and carbonic acid are housed
Lithium 0.09-0.11g；
No. 3 bottles: clarifier 95-105ml is housed；
No. 4 bottles: equipped with the neutral Instant cement 95-105ml for moist mounting.
In headpin, the component following containing parts by weight in Harri hematoxylin dye liquor: 1 part of hematoxylin, dehydrated alcohol 7.5-
8.5 parts, 19-21 parts of aluminum aluminum sulfate, 190-210 parts of distilled water, 0.45-0.55 parts of mercury oxide, 6-8 parts of glacial acetic acid；It prepares
When, hematoxylin is dissolved with dehydrated alcohol, dissolves by heating aluminum aluminum sulfate with distilled water, then two liquid merging is boiled, oxygen is added
Change mercury, continues heating and stirring solution to darkviolet, it is cooling with ice water immediately, restore to room temperature to filter spare, adds ice
Acetic acid simultaneously mixes, filters acquisition Harri hematoxylin dye liquor；Contain the component that parts by weight are following in ClarkeShi fixer: anhydrous
55-65 parts of ethyl alcohol, 18-22 parts of glacial acetic acid；Solution preparation process is to take Harri hematoxylin dye liquor in headpin, slow with glass rod
The ClarkeShi fixer that corresponding amount is gradually poured into while stirring, stands after being sufficiently stirred, and bottles after 3-8 minutes.
The configuration flow of solution is in No. 2 bottles: spirit soluble eosin Y being poured into dehydrated alcohol, glass rod while pouring into
Stirring is poured into distilled water after standing 2-4 minutes and is stirred with glass rod, is subsequently poured into lithium carbonate powder, glass rod while pouring into
Stirring, stands after being sufficiently stirred, bottles after 3-8 minutes.
Component following containing parts by weight in clarifier in No. 3 bottles: 20-60 parts of white oil, 12-45 parts of nonane, octane 6-35
Part, 5-20 parts of heptane, 4-25 parts of castor oil, 3-18 parts of amylalcohol and 2-9 parts of hexanol.
Neutral Instant cement in No. 4 bottles is " river original board " neutral quick-drying of Wuxi Jiangyuan Industry Company Jiangsu Institute of Nuclear Medicine's production
Glue.
Wherein, the ClarkeShi fixer in headpin can play the role of sloughing grease, protect nucleic acid.Dehydrated alcohol can
By the free moisture removal in tissue, albumen hydrophobic region moves on to protein surface, destroys the three-dimensional structure of protein.Dehydrated alcohol
It is for fixing simultaneously, also act the effect of dehydration；Glacial acetic acid solidifiable intracellular nucleic acid is ionized by changing
The charge of protein side chains, such as (- NH2-→NH3+) and (COO → COOH), it destroys electrostatic and is connect with chloro, by the anion of lipophilic
It is inserted into hydrophilic area, plays the role of the three-dimensional structure for destroying protein.Glacial acetic acid it is for fixing simultaneously, additionally it is possible to it is logical
Crossing the acidity of itself makes the pH < 5.0 of Harri hematoxylin dye liquor, colours cytoplasm not in such circumstances, and nucleus
Dye situation did not occurred, Harri hematoxylin dye liquor is promoted to colour the satisfied of nucleus.
Dehydrated alcohol in No. 2 bottles plays the role of continuing dehydration, and matches according to reagent, concentration of alcohol in No. 2 bottles
About 92%, substantially meet requirement of the ClarkeShi fixer to subsequent ethanol concentration in headpin；Lithium carbonate is dissolved in distilled water
Another entire solution also plays differentiation in this way and while subacidity reagent plays neutralization in headpin in alkalescence afterwards
Blue effect is returned, promotes spirit soluble eosin Y to cytoplasmic red dye.
It is dissolved in the corresponding clarifier of No. 3 bottles through remaining ethanol reagent on headpin and No. 2 bottles treated slide,
And it can be rinsed, while the effective ingredient in clarifier is woven with transparent, fixed effect to group, reaches during use
Continue transparent, fixed treatment effect to tissue, and without containing volatile hazardous substances such as dimethylbenzene, acetone.
The neutral Instant cement of No. 4 bottles can be changed in slide there are moist mounting is achieved the effect that in the case where clarifier
The deficiency for needing to blot mounting after residual solution on slide during existing film-making, when having saved film-making using this mounting mode
Between, it does not need to need to dry as existing film-making process mounting after tissue and expends the time, while not containing volatile injurant
Matter has no adverse effect to environment and film-making personnel.
Above-mentioned pathological diagnosis can be used for quickly handling a plurality of types of pathological diagnosis tissues with kit, and the present invention is also
A kind of immediate processing method of pathological diagnosis tissue is provided, tissue is carried out using above-mentioned 1,2,3, No. 4 washing bottle
It handles and in turn includes the following steps:
1) headpin stands 65-125s after dripping full slide；
2) with the micro- flushing of No. 2 bottles, (micro- flushing, which refers to, is rinsed the tissue on slide, and not after solution on abandoning slide
Tissue is set to fall off, shift, similarly hereinafter), until on slide without blue display；
3) 5-25s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean；
5) mounting after No. 4 bottle dropping liquid 0.1ml.It is required according to the film-making of different type pathological diagnosis tissue, this step
It can be omitted when making some types of tissue pieces.
According to handled different type pathological diagnosis tissue, it is segmented into following several situations and is handled:
First, handled tissue is superficial or Deep Lesions scraping blade or smear, the superficial or Deep Lesions scraping blade
Or smear includes at least cervical smear, sputum smear, urine smear, prostatic fluid smear, body surface secretion smear, needle and is emptied chamber liquid
One of body smear,
Processing step is successively are as follows:
1) headpin stands 70 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 10 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean.
Second, handled tissue is aspiration smear, direct view under endoscope smear or mammary gland discharge smear under image check,
Step is successively are as follows:
1) headpin stands 90 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 15 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean；
5) mounting after No. 4 bottle dropping liquid 0.1ml.
Aspiration smear, direct view under endoscope smear or nipple discharge smear include at least first shape under B ultrasound under the image check
Nylon bruss smear and nipple hemorrhage apply under gland puncture smear, B ultrasound inferior gluteal lymph node aspiration smear, CT Conducted Puncture smear, gastroscope
One of piece.
Third, handled tissue is quick frozen-section, step is successively are as follows:
1) headpin stands 120 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 20 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean；
5) mounting after No. 4 bottle dropping liquid 0.1ml.
Compared with prior art, what the present invention obtained has the beneficial effect that
The time needed for 1. existing tissue processing is greatly reduced.The dozens of minutes as needed for existing processing is to a few hours
It differs, is reduced to complete in several minutes, improve work efficiency.
2. considerably reducing the process links of existing tissue processing.Process by significantly reducing existing tissue processing
Link also reduces the generation of miswork.
3. reducing the consumption of various reagents, environmental pollution is reduced.By centralized processing needed for the processing of existing tissue
It is changed into slide and dyeing is added dropwise, reduces the usage amount of reagent, improve the utilization rate of reagent.Existing tissue is given up simultaneously
The volatile hazardous substances such as dimethylbenzene, acetone in processing, reduce environmental pollution.
4. false positive results caused by avoiding because of cross contamination in the processing of existing tissue.Because of the processing of existing tissue
It is middle by the way of centralized processing, which results in may because on positive slide lesion ingredient fall off caused by other negative slides
Pollution.And this technology takes every slide that facture is added dropwise, and avoids the mutual pollution between inspection slide.
5. requirement of the tissue processing to space is reduced, conducive to the development to medical services from far-off regions.Existing tissue
Because reagent is numerous in piece treatment process, more space is occupied, is unfavorable for carrying.In volunteer medical consultation from far-off regions, generaI investigation mostly by clinic
Tissue is taken back post-processing by doctor, and pathological diagnosis result is fed back to locality after a few days；And required reagent is placed in 4 by this technology
It is easy to carry in a washing bottle, it can accomplish to check have pathology diagnostic result on the spot on the spot in volunteer medical consultation from far-off regions, generaI investigation, benefit
In the raising of medical service level from far-off regions.
Detailed description of the invention
Fig. 1 is cervical smear treated image, 10 × 10 times of amplification factor.
Fig. 2 is hydrothorax smear treated image, 10 × 10 times of amplification factor.
Fig. 3 is nipple discharge smear treated image, 10 × 20 times of amplification factor.
Fig. 4 is B ultrasound thyroid aspiration smear treated image, 10 × 10 times of amplification factor.
Fig. 5 is quick frozen-section treated image, 10 × 10 times of amplification factor.
Specific embodiment
Embodiment 1
A kind of pathological diagnosis pathological diagnosis kit is provided with four washing bottles with identification label, marks respectively
It is denoted as headpin, No. 2 bottles, No. 3 bottles and No. 4 bottles, wherein
Headpin: Harri hematoxylin dye liquor 85ml and ClarkeShi fixer 42ml is housed；
In headpin, the component following containing parts by weight in Harri hematoxylin dye liquor: 1 part of hematoxylin, dehydrated alcohol 7.5
Part, 19 parts of aluminum aluminum sulfate, 190 parts of distilled water, 0.55 part of mercury oxide, 6 parts of glacial acetic acid；When preparation, dissolved with dehydrated alcohol
Hematoxylin dissolves by heating aluminum aluminum sulfate with distilled water, then boils two liquid merging, mercury oxide is added, continues to heat and stir
Solution is mixed to darkviolet, it is cooling with ice water immediately, restore to room temperature to filter spare, adds glacial acetic acid and mix, filter and obtain
Obtain Harri hematoxylin dye liquor；
Contain the component that parts by weight are following: dehydrated alcohol 60ml, glacial acetic acid 20ml in ClarkeShi fixer；
Solution preparation process is in headpin: taking Harri hematoxylin dye liquor, gradually pours into while being slowly stirred with glass rod
The ClarkeShi fixer of corresponding amount, stands after being sufficiently stirred, bottles after five minutes.
No. 2 bottles: spirit soluble eosin Y 0.38g, dehydrated alcohol 105ml, distilled water 12ml and lithium carbonate 0.09g are housed；Its
Middle eosin Y is produced by Sinopharm Chemical Reagent Co., Ltd., lot number F20100302；
The configuration flow of solution is in No. 2 bottles: spirit soluble eosin Y being poured into dehydrated alcohol, glass rod while pouring into
Stirring is poured into distilled water after standing 3 minutes and is stirred with glass rod, is subsequently poured into lithium carbonate powder, stirred while pouring into glass rod
It mixes, stands after being sufficiently stirred, bottle after five minutes.
No. 3 bottles: clarifier 105ml is housed；
Component following containing parts by weight in clarifier in No. 3 bottles: 20 parts of white oil, 45 parts of nonane, 6 parts of octane, heptane 5
Part, 25 parts of castor oil, 3 parts of amylalcohol and 9 parts of hexanol.
No. 4 bottles: equipped with the neutral Instant cement 105ml for moist mounting.
Neutral Instant cement in No. 4 bottles is " river original board " neutral quick-drying of Wuxi Jiangyuan Industry Company Jiangsu Institute of Nuclear Medicine's production
Glue, product batch number 14050809.
Wherein, the ClarkeShi fixer in headpin can play the role of sloughing grease, protect nucleic acid.Dehydrated alcohol can
By the free moisture removal in tissue, albumen hydrophobic region moves on to protein surface, destroys the three-dimensional structure of protein.Dehydrated alcohol
It is for fixing simultaneously, also act the effect of dehydration；Glacial acetic acid solidifiable intracellular nucleic acid is ionized by changing
The charge of protein side chains, such as (- NH2-→NH3+) and (COO → COOH), it destroys electrostatic and is connect with chloro, by the anion of lipophilic
It is inserted into hydrophilic area, plays the role of the three-dimensional structure for destroying protein.Glacial acetic acid it is for fixing simultaneously, additionally it is possible to it is logical
Crossing the acidity of itself makes the pH < 5.0 of Harri hematoxylin dye liquor, colours cytoplasm not in such circumstances, and nucleus
Dye situation did not occurred, Harri hematoxylin dye liquor is promoted to colour the satisfied of nucleus.
Dehydrated alcohol in No. 2 bottles plays the role of continuing dehydration, and matches according to reagent, concentration of alcohol in No. 2 bottles
About 92%, substantially meet requirement of the ClarkeShi fixer to subsequent ethanol concentration in headpin；Lithium carbonate is dissolved in distilled water
Another entire solution also plays differentiation in this way and while subacidity reagent plays neutralization in headpin in alkalescence afterwards
Blue effect is returned, promotes spirit soluble eosin Y to cytoplasmic red dye.
Through headpin, remaining ethanol reagent is dissolved in the corresponding clarifier of No. 3 bottles on slide after No. 2 bottles, and can quilt
It rinses out, while the effective ingredient in clarifier is woven with transparent, fixed effect to group, has reached during use to group
It knits and continues transparent, fixed treatment effect, and without containing volatile hazardous substances such as dimethylbenzene, acetone.
The neutral Instant cement of No. 4 bottles can be changed in slide there are moist mounting is achieved the effect that in the case where clarifier
The deficiency for needing to blot mounting after residual solution on slide during existing film-making, when having saved film-making using this mounting mode
Between, it does not need to need to dry as existing film-making process mounting after tissue and expends the time, while not containing volatile injurant
Matter has no adverse effect to environment and film-making personnel.
Embodiment 2
A kind of pathological diagnosis pathological diagnosis kit is provided with four washing bottles with identification label, marks respectively
It is denoted as headpin, No. 2 bottles, No. 3 bottles and No. 4 bottles, wherein
Headpin: Harri hematoxylin dye liquor 95ml and ClarkeShi fixer 38ml is housed；
In headpin, the component following containing parts by weight in Harri hematoxylin dye liquor: 1 part of hematoxylin, dehydrated alcohol 8.5
Part, 20 parts of aluminum aluminum sulfate, 200 parts of distilled water, 0. 5 parts of mercury oxide, 8 parts of glacial acetic acid；When preparation, is dissolved and revived with dehydrated alcohol
Another name for dissolves by heating aluminum aluminum sulfate with distilled water, then boils two liquid merging, mercury oxide is added, continue heating and stirring
Solution is cooling with ice water immediately to darkviolet, restores to room temperature to filter spare, adds glacial acetic acid and mixes, filters acquisition
Harri hematoxylin dye liquor；
Contain the component that parts by weight are following in ClarkeShi fixer: 55 parts of dehydrated alcohol, 22 parts of glacial acetic acid；
Solution preparation process is in headpin: taking Harri hematoxylin dye liquor, gradually pours into while being slowly stirred with glass rod
The ClarkeShi fixer of corresponding amount, stands after being sufficiently stirred, and bottles after 3 minutes.
No. 2 bottles: spirit soluble eosin Y 0.42g, dehydrated alcohol 115ml, distilled water 9ml and lithium carbonate 0.11g are housed；
The configuration flow of solution is in No. 2 bottles: spirit soluble eosin Y being poured into dehydrated alcohol, glass rod while pouring into
Stirring is poured into distilled water after standing 2 minutes and is stirred with glass rod, is subsequently poured into lithium carbonate powder, stirred while pouring into glass rod
It mixes, is stood after being sufficiently stirred, bottled after 8 minutes.
No. 3 bottles: clarifier 95ml is housed；
Component following containing parts by weight in clarifier in No. 3 bottles: 60 parts of white oil, 12 parts of nonane, 35 parts of octane, heptane 20
Part, 4 parts of castor oil, 18 parts of amylalcohol and 2 parts of hexanol.
No. 4 bottles: equipped with the neutral Instant cement 95ml for moist mounting.
Neutral Instant cement in No. 4 bottles is " river original board " neutral quick-drying of Wuxi Jiangyuan Industry Company Jiangsu Institute of Nuclear Medicine's production
Glue, product batch number 14050809.
Embodiment 3
A kind of pathological diagnosis pathological diagnosis kit is provided with four washing bottles with identification label, marks respectively
It is denoted as headpin, No. 2 bottles, No. 3 bottles and No. 4 bottles, wherein
Headpin: Harri hematoxylin dye liquor 90ml and ClarkeShi fixer 40ml is housed；
In headpin, the component following containing parts by weight in Harri hematoxylin dye liquor: 1 part of hematoxylin, 8 parts of dehydrated alcohol,
21 parts of aluminum aluminum sulfate, 210 parts of distilled water, 0.45 part of mercury oxide, 6.5 parts of glacial acetic acid；When preparation, is dissolved and revived with dehydrated alcohol
Another name for dissolves by heating aluminum aluminum sulfate with distilled water, then boils two liquid merging, mercury oxide is added, continue heating and stirring
Solution is cooling with ice water immediately to darkviolet, restores to room temperature to filter spare, adds glacial acetic acid and mixes, filters acquisition
Harri hematoxylin dye liquor；
Contain the component that parts by weight are following in ClarkeShi fixer: 65 parts of dehydrated alcohol, 18 parts of glacial acetic acid；
Solution preparation process is in headpin: taking Harri hematoxylin dye liquor, gradually pours into while being slowly stirred with glass rod
The ClarkeShi fixer of corresponding amount, stands after being sufficiently stirred, and bottles after 8 minutes.
No. 2 bottles: spirit soluble eosin Y 0.4g, dehydrated alcohol 110ml, distilled water 10ml and lithium carbonate 0.1g are housed；
The configuration flow of solution is in No. 2 bottles: spirit soluble eosin Y being poured into dehydrated alcohol, glass rod while pouring into
Stirring is poured into distilled water after standing 4 minutes and is stirred with glass rod, is subsequently poured into lithium carbonate powder, stirred while pouring into glass rod
It mixes, is stood after being sufficiently stirred, bottled after 8 minutes.
No. 3 bottles: clarifier 100ml is housed；
Component following containing parts by weight in clarifier in No. 3 bottles: 40 parts of white oil, 30 parts of nonane, 25 parts of octane, heptane 15
Part, 15 parts of castor oil, 12 parts of amylalcohol and 6 parts of hexanol.
No. 4 bottles: equipped with the neutral Instant cement 100ml for moist mounting.
Neutral Instant cement in No. 4 bottles is " river original board " neutral quick-drying of Wuxi Jiangyuan Industry Company Jiangsu Institute of Nuclear Medicine's production
Glue, product batch number 14050809.
Embodiment 4
A kind of pathological diagnosis pathological diagnosis kit is provided with four washing bottles with identification label, marks respectively
It is denoted as headpin, No. 2 bottles, No. 3 bottles and No. 4 bottles, wherein
Headpin: Harri hematoxylin dye liquor 85ml and ClarkeShi fixer 40ml is housed；
In headpin, the component following containing parts by weight in Harri hematoxylin dye liquor: 1 part of hematoxylin, dehydrated alcohol 8.5
Part, 21 parts of aluminum aluminum sulfate, 190 parts of distilled water, 0.455 part of mercury oxide, 8 parts of glacial acetic acid；When preparation, dissolved with dehydrated alcohol
Hematoxylin dissolves by heating aluminum aluminum sulfate with distilled water, then boils two liquid merging, mercury oxide is added, continues to heat and stir
Solution is mixed to darkviolet, it is cooling with ice water immediately, restore to room temperature to filter spare, adds glacial acetic acid and mix, filter and obtain
Obtain Harri hematoxylin dye liquor；
Contain the component that parts by weight are following in ClarkeShi fixer: 55 parts of dehydrated alcohol, 18 parts of glacial acetic acid；
Solution preparation process is in headpin: taking Harri hematoxylin dye liquor, gradually pours into while being slowly stirred with glass rod
The ClarkeShi fixer of corresponding amount, stands after being sufficiently stirred, and bottles after 3 minutes.
No. 2 bottles: spirit soluble eosin Y 0.42g, dehydrated alcohol 115ml, distilled water 12ml and lithium carbonate 0.09g are housed；
The configuration flow of solution is in No. 2 bottles: spirit soluble eosin Y being poured into dehydrated alcohol, glass rod while pouring into
Stirring is poured into distilled water after standing 4 minutes and is stirred with glass rod, is subsequently poured into lithium carbonate powder, stirred while pouring into glass rod
It mixes, is stood after being sufficiently stirred, bottled after 8 minutes.
No. 3 bottles: clarifier 95ml is housed；
Component following containing parts by weight in clarifier in No. 3 bottles: 60 parts of white oil, 125 parts of nonane, 35 parts of octane, heptane 5
Part, 25 parts of castor oil, 3 parts of amylalcohol and 9 parts of hexanol.
No. 4 bottles: equipped with the neutral Instant cement 95ml for moist mounting.
Neutral Instant cement in No. 4 bottles is " river original board " neutral quick-drying of Wuxi Jiangyuan Industry Company Jiangsu Institute of Nuclear Medicine's production
Glue.
Embodiment 5
A kind of pathological diagnosis pathological diagnosis kit is provided with four washing bottles with identification label, marks respectively
It is denoted as headpin, No. 2 bottles, No. 3 bottles and No. 4 bottles, wherein
Headpin: Harri hematoxylin dye liquor 95ml and ClarkeShi fixer 40ml is housed；
In headpin, the component following containing parts by weight in Harri hematoxylin dye liquor: 1 part of hematoxylin, 8 parts of dehydrated alcohol,
20 parts of aluminum aluminum sulfate, 200 parts of distilled water, 0.45 part of mercury oxide, 6 parts of glacial acetic acid；When preparation, bush is dissolved with dehydrated alcohol
Essence dissolves by heating aluminum aluminum sulfate with distilled water, then boils two liquid merging, mercury oxide is added, it is molten to continue heating and stirring
Liquid is cooling with ice water immediately to darkviolet, restores to room temperature to filter spare, adds glacial acetic acid and mixes, filters acquisition
Harri hematoxylin dye liquor；
Contain the component that parts by weight are following in ClarkeShi fixer: 65 parts of dehydrated alcohol, 22 parts of glacial acetic acid；
Solution preparation process is in headpin: taking Harri hematoxylin dye liquor, gradually pours into while being slowly stirred with glass rod
The ClarkeShi fixer of corresponding amount, stands after being sufficiently stirred, bottles after five minutes.
No. 2 bottles: spirit soluble eosin Y 0.4g, dehydrated alcohol 110ml, distilled water 11ml and lithium carbonate 0.11g are housed；
The configuration flow of solution is in No. 2 bottles: spirit soluble eosin Y being poured into dehydrated alcohol, glass rod while pouring into
Stirring is poured into distilled water after standing 2 minutes and is stirred with glass rod, is subsequently poured into lithium carbonate powder, stirred while pouring into glass rod
It mixes, is stood after being sufficiently stirred, bottled after 3 minutes.
No. 3 bottles: clarifier 105ml is housed；
Component following containing parts by weight in clarifier in No. 3 bottles: 300 parts of white oil, 12 parts of nonane, 35 parts of octane, heptane
20 parts, 4 parts of castor oil, 5 parts of amylalcohol and 2 parts of hexanol.
No. 4 bottles: equipped with the neutral Instant cement 95ml for moist mounting.
Neutral Instant cement in No. 4 bottles is " river original board " neutral quick-drying of Wuxi Jiangyuan Industry Company Jiangsu Institute of Nuclear Medicine's production
Glue.
Embodiment 6
The pathological diagnosis of above-described embodiment 1-5 can be used for quickly handling a plurality of types of pathological diagnosis tissues with kit
Piece is in turn included the following steps when being handled using above-mentioned 1,2,3, No. 4 washing bottle tissue:
1) headpin stands 65-125s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 5-25s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean.
5) mounting after No. 4 bottle dropping liquid 0.1ml.
Wherein, step 5) can be required according to the film-making of different type pathological diagnosis tissue, make some type groups
It can be omitted when knitting piece.
Embodiment 7
As shown in Figure 1, handling cervical smear with the kit of above-described embodiment 1, step is successively are as follows:
1) headpin stands 70 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 10 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean.
This method can be handled for handling superficial or Deep Lesions scraping blade or smear including cervical smear, sputum smear, urine
Smear, prostatic fluid smear, body surface secretion smear, needle inhale cavity liquid smear etc..
As shown in Fig. 2, for treated, needle inhales cavity liquid smear, specially hydrothorax smear.
The present embodiment is sufficiently with the taken tissue superficial of such tissue, characteristic that is loose and being easy to coloring, headpin and 2
Number bottle processing time is most short, and handles without mounting to reduce the processing time, can show karyon indigo plant dye, the red dye of cytoplasm, and right
Than clear, reach the quality requirement of pathological diagnosis.
Embodiment 8
As shown in figure 3, handling nipple hemorrhage smear with the kit of above-described embodiment 2, step is successively are as follows:
1) headpin stands 90 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 15 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean；
5) mounting after No. 4 bottle dropping liquid 0.1ml.
Aspiration smear under image check, direct view under endoscope smear, including B ultrasound thyroid aspiration smear, B can also be handled
Nylon bruss smear etc. under super inferior gluteal lymph node aspiration smear, CT Conducted Puncture smear, gastroscope.
As shown in figure 4, for the picture of treated B ultrasound thyroid aspiration smear.
Adhesion strength strong, headpin and 2 of comparing adhesion strength between scraping blade and smear tissue between the taken tissue of the tissue of the present embodiment
Number bottle processing time extends, and the refractivity of tissue is improved after needing mounting to handle, and karyon can be shown after above-mentioned processing
Indigo plant dye, the red dye of cytoplasm, and compare clearly, reach the quality requirement of pathological diagnosis.
Embodiment 9
As shown in figure 5, step is successively with the kit quick frozen-section of above-described embodiment 3 are as follows:
1) headpin stands 120 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 20 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean；
5) mounting after No. 4 bottle dropping liquid 0.1ml.
Adhesion strength is most strong between the taken tissue of the tissue of the present embodiment, thus headpin and No. 2 bottles handle time longest, and
The refractivity that tissue is improved after needing mounting to handle can show karyon indigo plant dye, the red dye of cytoplasm after above-mentioned processing, and compare
Clearly, reach the quality requirement of pathological diagnosis.
The present invention is not limited to the above embodiments, on the basis of technical solution disclosed by the invention, the skill of this field
For art personnel according to disclosed technology contents, one can be made to some of which technical characteristic by not needing creative labor
A little replacements and deformation, these replacements and deformation are within the scope of the invention.
Claims (10)
1. a kind of pathological diagnosis tissue treatment kits, which is characterized in that four washing bottles including having identification label,
Wherein,
Headpin: Harri hematoxylin dye liquor 85-95ml and ClarkeShi fixer 38-42ml is housed；
No. 2 bottles: spirit soluble eosin Y 0.38-0.42g, dehydrated alcohol 105-115ml, distilled water 9-12ml and lithium carbonate are housed
0.09-0.11g；
No. 3 bottles: clarifier 95-105ml is housed；
No. 4 bottles: equipped with the neutral Instant cement 95-105ml for moist mounting.
2. a kind of pathological diagnosis tissue treatment kits according to claim 1, which is characterized in that in headpin,
The component following containing parts by weight in Harri hematoxylin dye liquor:
1 part of hematoxylin, 7.5-8.5 parts of dehydrated alcohol, 19-21 parts of aluminum aluminum sulfate, 190-210 parts of distilled water, mercury oxide
0.45-0.55 parts, 6-8 parts of glacial acetic acid；
When preparation, hematoxylin is dissolved with dehydrated alcohol, dissolves by heating aluminum aluminum sulfate with distilled water, then boils two liquid merging
Mercury oxide is added in boiling, continues heating and stirring solution to darkviolet, cooling with ice water immediately, restore to room temperature to filter it is spare,
It adds glacial acetic acid and mixes, filters acquisition Harri hematoxylin dye liquor；
Contain the component that parts by weight are following in ClarkeShi fixer: 55-65 parts of dehydrated alcohol, 18-22 parts of glacial acetic acid；
Solution preparation process is to take Harri hematoxylin dye liquor in headpin, is gradually poured into while being slowly stirred with glass rod corresponding
The ClarkeShi fixer of amount, stands after being sufficiently stirred, and bottles after 3-8 minutes.
3. a kind of pathological diagnosis tissue treatment kits according to claim 1, which is characterized in that molten in No. 2 bottles
The configuration flow of liquid is: spirit soluble eosin Y being poured into dehydrated alcohol, is stirred while pouring into glass rod, after standing 2-4 minutes
It pours into distilled water and is stirred with glass rod, be subsequently poured into lithium carbonate powder, stirred while pouring into glass rod, be sufficiently stirred rear quiet
It sets, bottles after 3-8 minutes.
4. a kind of pathological diagnosis tissue treatment kits according to claim 1, which is characterized in that in No. 3 bottles thoroughly
Contain the component that parts by weight are following: 20-60 parts of white oil, 12-45 parts of nonane, 6-35 parts of octane, 5-20 parts of heptane, castor-oil plant in bright dose
4-25 parts oily, 3-18 parts of amylalcohol and 2-9 parts of hexanol.
5. a kind of pathological diagnosis tissue treatment kits according to claim 1, which is characterized in that in No. 4 bottles
Neutral Instant cement is " river original board " neutral Instant cement of Wuxi Jiangyuan Industry Company Jiangsu Institute of Nuclear Medicine's production.
6. a kind of immediate processing method of pathological diagnosis tissue, which is characterized in that use the 1 of claim 1,2,3, No. 4
Washing bottle successively follows the steps below processing with tissue to different types of pathological diagnosis:
1) headpin stands 65-125s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 5-25s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean.
7. a kind of immediate processing method of pathological diagnosis tissue according to claim 6, which is characterized in that handled
Tissue be that superficial or Deep Lesions scraping blade or smear, the superficial or Deep Lesions scraping blade or smear are scraped including at least uterine neck
Piece, sputum smear, urine smear, prostatic fluid smear, body surface secretion smear, needle inhale one of cavity liquid smear, specific steps
Successively are as follows:
1) headpin stands 70 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 10 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean.
8. a kind of immediate processing method of pathological diagnosis tissue according to claim 6, which is characterized in that in step
It 4) further include mounting after No. 4 bottle dropping liquids of step 5) after.
9. a kind of immediate processing method of pathological diagnosis tissue according to claim 8, which is characterized in that handled
Tissue be image check under aspiration smear, direct view under endoscope smear or nipple discharge smear, step is successively are as follows:
1) headpin stands 90 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 15 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean；
5) mounting after No. 4 bottle dropping liquid 0.1ml.
10. a kind of immediate processing method of pathological diagnosis tissue according to claim 8, which is characterized in that locating
The tissue of reason is quick frozen-section, and step is successively are as follows:
1) headpin stands 120 ± 5s after dripping full slide；
2) it abandons and uses No. 2 micro- flushings of bottle on slide after solution, until being shown on slide without blue；
3) 20 ± 5s is stood after dripping full slide with No. 2 bottles；
4) it abandons on slide after solution with the micro- flushing slide of No. 3 bottles to clean；
5) mounting after No. 4 bottle dropping liquid 0.1ml.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710651672.2A CN107490506B (en) | 2017-08-02 | 2017-08-02 | The immediate processing method of pathological diagnosis kit and tissue |
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| US6203394B1 (en) * | 2000-01-07 | 2001-03-20 | Vincent Kuo Wei Lee | Ornamental liquid color box |
| CN101344467A (en) * | 2008-08-20 | 2009-01-14 | 赵晓辉 | Routine paraffin wax flaking HE dyeing non-xylol whole-course treatment liquid for biological organization sample |
| CN102147337A (en) * | 2011-01-13 | 2011-08-10 | 中国水产科学研究院东海水产研究所 | Paraffin sectioning method for ocean shellfish D-shaped larva |
| CN102876081A (en) * | 2012-09-24 | 2013-01-16 | 广州鸿琪光学仪器科技有限公司 | Hematoxylin staining solution and papanicolaou staining kit containing same |
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| US6203394B1 (en) * | 2000-01-07 | 2001-03-20 | Vincent Kuo Wei Lee | Ornamental liquid color box |
| CN101344467A (en) * | 2008-08-20 | 2009-01-14 | 赵晓辉 | Routine paraffin wax flaking HE dyeing non-xylol whole-course treatment liquid for biological organization sample |
| CN102147337A (en) * | 2011-01-13 | 2011-08-10 | 中国水产科学研究院东海水产研究所 | Paraffin sectioning method for ocean shellfish D-shaped larva |
| CN102876081A (en) * | 2012-09-24 | 2013-01-16 | 广州鸿琪光学仪器科技有限公司 | Hematoxylin staining solution and papanicolaou staining kit containing same |
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