CN106442074A - Reticular fiber staining kit for pathological examination - Google Patents
Reticular fiber staining kit for pathological examination Download PDFInfo
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- CN106442074A CN106442074A CN201610740723.4A CN201610740723A CN106442074A CN 106442074 A CN106442074 A CN 106442074A CN 201610740723 A CN201610740723 A CN 201610740723A CN 106442074 A CN106442074 A CN 106442074A
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- 230000001575 pathological effect Effects 0.000 title claims abstract description 30
- 238000010186 staining Methods 0.000 title claims abstract description 25
- 108010081750 Reticulin Proteins 0.000 title claims abstract description 23
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 53
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 claims abstract description 42
- 238000000034 method Methods 0.000 claims abstract description 28
- 238000005406 washing Methods 0.000 claims abstract description 26
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 claims abstract description 20
- 235000019345 sodium thiosulphate Nutrition 0.000 claims abstract description 20
- LCPUDZUWZDSKMX-UHFFFAOYSA-K azane;hydrogen sulfate;iron(3+);sulfate;dodecahydrate Chemical compound [NH4+].O.O.O.O.O.O.O.O.O.O.O.O.[Fe+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O LCPUDZUWZDSKMX-UHFFFAOYSA-K 0.000 claims abstract description 16
- 235000006408 oxalic acid Nutrition 0.000 claims abstract description 14
- 239000000243 solution Substances 0.000 claims abstract description 13
- FDWREHZXQUYJFJ-UHFFFAOYSA-M gold monochloride Chemical compound [Cl-].[Au+] FDWREHZXQUYJFJ-UHFFFAOYSA-M 0.000 claims abstract description 12
- 239000007864 aqueous solution Substances 0.000 claims abstract description 11
- 239000012286 potassium permanganate Substances 0.000 claims abstract description 5
- -1 diamine silver hydroxide Chemical class 0.000 claims abstract 5
- 238000004821 distillation Methods 0.000 claims description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 23
- 238000004043 dyeing Methods 0.000 claims description 23
- 239000000835 fiber Substances 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 239000012153 distilled water Substances 0.000 claims description 15
- NCPUOHFQDGQSAS-UHFFFAOYSA-M N[Ag](N)O Chemical compound N[Ag](N)O NCPUOHFQDGQSAS-UHFFFAOYSA-M 0.000 claims description 10
- 230000018044 dehydration Effects 0.000 claims description 6
- 238000006297 dehydration reaction Methods 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 6
- 238000004061 bleaching Methods 0.000 claims description 5
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- 239000005864 Sulphur Substances 0.000 claims 1
- 229920000642 polymer Polymers 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- 230000007170 pathology Effects 0.000 abstract description 11
- 208000015914 Non-Hodgkin lymphomas Diseases 0.000 abstract description 7
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 abstract description 6
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 102000004169 proteins and genes Human genes 0.000 abstract description 6
- GGCZERPQGJTIQP-UHFFFAOYSA-N sodium;9,10-dioxoanthracene-2-sulfonic acid Chemical compound [Na+].C1=CC=C2C(=O)C3=CC(S(=O)(=O)O)=CC=C3C(=O)C2=C1 GGCZERPQGJTIQP-UHFFFAOYSA-N 0.000 abstract description 6
- 238000001514 detection method Methods 0.000 abstract description 5
- 208000029922 reticulum cell sarcoma Diseases 0.000 abstract description 4
- 229910052709 silver Inorganic materials 0.000 abstract description 4
- 239000004332 silver Substances 0.000 abstract description 4
- 206010028980 Neoplasm Diseases 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 abstract 1
- 201000004477 skin sarcoma Diseases 0.000 abstract 1
- 210000001519 tissue Anatomy 0.000 description 30
- 239000007788 liquid Substances 0.000 description 5
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000003086 colorant Substances 0.000 description 4
- 238000005034 decoration Methods 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 206010012818 diffuse large B-cell lymphoma Diseases 0.000 description 3
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 3
- 239000010931 gold Substances 0.000 description 3
- 229910052737 gold Inorganic materials 0.000 description 3
- 238000005213 imbibition Methods 0.000 description 3
- 201000006845 reticulosarcoma Diseases 0.000 description 3
- 244000025254 Cannabis sativa Species 0.000 description 2
- 238000005660 chlorination reaction Methods 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 208000025036 lymphosarcoma Diseases 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- UHOVQNZJYSORNB-UHFFFAOYSA-N monobenzene Natural products C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- NDVLTYZPCACLMA-UHFFFAOYSA-N silver oxide Chemical compound [O-2].[Ag+].[Ag+] NDVLTYZPCACLMA-UHFFFAOYSA-N 0.000 description 2
- 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 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 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 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- SJEYSFABYSGQBG-UHFFFAOYSA-M Patent blue Chemical compound [Na+].C1=CC(N(CC)CC)=CC=C1C(C=1C(=CC(=CC=1)S([O-])(=O)=O)S([O-])(=O)=O)=C1C=CC(=[N+](CC)CC)C=C1 SJEYSFABYSGQBG-UHFFFAOYSA-M 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- SNDJGKIVHKOEHY-UHFFFAOYSA-M S(=S)(=O)(O)O.S[Na] Chemical compound S(=S)(=O)(O)O.S[Na] SNDJGKIVHKOEHY-UHFFFAOYSA-M 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- FOIXSVOLVBLSDH-UHFFFAOYSA-N Silver ion Chemical compound [Ag+] FOIXSVOLVBLSDH-UHFFFAOYSA-N 0.000 description 1
- 239000000980 acid dye Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000001465 metallisation Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229910001923 silver oxide Inorganic materials 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000010998 test method 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
-
- 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
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 reticular fiber staining kit for pathological examination, wherein the kit mainly includes potassium permanganate, oxalic acid, iron alum, diamine silver hydroxide, formaldehyde, gold chloride and sodium thiosulfate; the principle includes that silver ions in a diamine silver hydroxide aqueous solution are combined with proteins in a stained tissue, and black metallic silver formed by reducing the silver ions with formaldehyde is deposited in the tissue and on the tissue surface; after gold chloride is used for adjusting a color, then a sodium thiosulfate solution is used for washing unreduced silver salt, and reticular fibers in the tissue are clearly displayed. The kit has the advantages of simple use method, easy operation, good staining effect, and little influence by external factors; reticular fiber staining is mainly applied for distinguishing properties and sources of tumor in pathology examination, and has wide application in hemangio skin sarcoma, lymphatic sarcoma, reticular cell sarcoma and other tumor detection fields.
Description
Technical field
The present invention relates to pathological examination technical field, more particularly to a kind of examination of reticular fiber staining for pathological examination
Agent box.
Background technology
Pathologic finding is a kind of Pathomorphology method with the pathological change in biological organs, tissue or cell.For visiting
Beg for the lysis that organ, tissue or cell are occurred, the method that can be checked using certain Pathomorphology, check that they are sent out
Raw pathology, inquires into pathology Producing reason, pathogenesis, the generation evolution of pathology, finally makes pathological diagnosis.Pathology
Morphologic inspection method, looks first at the pathological change of gross specimen, then cuts a certain size pathological tissues, uses pathology
Pathological section is made by Histological method, checks pathology further with microscope.Take a certain size pathological tissues, use pathological tissue
Method makes pathological section, checks pathology further with microscope.The generation evolution of pathology, finally makes pathology and examines
Disconnected.
Dyeing is a very crucial step in pathological section preparation process, and conventional colouring method is Hematoxylin-eosin dyeing
Method, abbreviation H.E decoration method.This method to the fixing tissue of any fixer and applies the section of various investments all can make
With.Haematoxylin is a kind of basic-dyeable fibre, and the basophilla material in tissue can be made to dye blueness, such as chromatin in nucleus etc.;
Yihong is a kind of acid dyes, and the acidophilia material in tissue can be made to dye redness, and the kytoplasm of such as most cells, kernel etc. exist
All take on a red color in the section of H.E dyeing.H.E dyeing procedure is as follows:Dewaxing, de- benzene, rehydration, dyeing, dehydration, transparent, sealing.And
For reticular fiber staining, due to reticular fibre, in loose connective tissue, content is less, and fiber is thinner, has branch, that
This intertexture reticulates, therefore inconspicuous using Hematoxylin-eosin decoration method Color, and in electric Microscopic observation, reticular fibre has
There is equidistant band structure, its chemical composition is also collagen, similar with collagenous fibres, sugared egg on collagenous fibres for the bag
Make reticular fibre have argyrophilia in vain, therefore soak silver-colored method and fiber can be dyed black, thus clearly presenting under Electronic Speculum
Come, this is a kind of new method of current reticular fiber staining.
Content of the invention
It is an object of the invention to provide a kind of reticular fiber staining kit for pathological examination.
The technical scheme is that:A kind of reticular fiber staining kit for pathological examination, main inclusion:Gao Meng
Sour potassium 5-10ml, oxalic acid 3-5ml, iron alum 3-5ml, diamino silver hydroxide 2-4ml, formaldehyde 1-3ml, chlorauride 5-10ml, sulphur
Sodium thiosulfate 2-5ml.
A kind of using method of the reticular fiber staining kit for pathological examination is:
A. the section preparing is carried out after dewaxing treatment, distilled water rinses 1-3 minute;
B. 2-4 minute, distillation washing 2-3 time are contaminated in liquor potassic permanganate;
C. after oxalic acid bleaching, distillation washing 2 times;
D. using iron alum aqueous solution mordant dyeing 5-10 minute, distillation washing 3-5 time;
E. diamino silver hydroxide contaminates 1-3 minute, distillation washing 3 times;
F. use formalin reductase 12-4 minutes, distilled water 1-3 time;
G. with chlorogold solution toning 3-5 minute, distilled water 2 times;
H. 3-5 minute, distillation washing 1-3 time are fixed with sodium thiosulfate;
I. use quality concentration is that 95% alcohol is suddenly washed, and until observing clearly till fiber, carries out weight when needing
Counterstain, carries out after the completion of dyeing being dehydrated, transparent and sealing.
Further, described liquor potassic permanganate mass concentration is 0.25-0.35%;Described oxalic acid mass concentration is
0.8-1%;Described iron alum aqueous solution mass concentration is 1.5-3.5%;Described formalin mass concentration is 8-12%;
Described chlorogold solution quality solubility is 0.2%;Described sodium thiosulfate mass concentration is 3-5%.
Further, after dyeing according to described using method, ammargenum is organized the protein combination in absorption and tissue,
It is deposited in tissue and surface through the argent that formaldehyde is reduced into black, after chlorauride toning, then washed with sodium thiosulfate liquid
Remove unreduced silver salt, so that in-house reticular fibre is clearly with black display out.
Further, described dehydration refers to carry out processed using absolute alcohol, and absolute alcohol has very strong suction
Aqueous, the hardening of tissue can be made again, make the colour developing of fibr tissue more stable.
The having the beneficial effect that of the kit of the present invention:For the netted fibre in loose connective tissue in pathological examination
Dimension hplc is less, and fiber is thinner, has branch, is interlaced with one another and reticulates, and has good coloration, in use diamino hydrogen
Protein combination in silver ion in the silver oxide aqueous solution and the tissue being colored, is reduced into the metal deposition of silver of black through formaldehyde
In tissue and surface, after being mixed colours with chlorauride, then wash away unreduced silver salt with sodium thiosulfate liquid, thus will be in-house
Reticular fibre clearly displays out, and the using method of kit of the present invention is simple, and easy to operate, Color is good, by extraneous because
Element impact is little, and reticular fiber staining is mainly used in property and the source of difference tumour in pathological examination, in blood vessel rind gall, pouring
The lesion detection such as bar sarcoma, reticulosarcoma field has and is widely applied very much.
Specific embodiment
Embodiment 1:A kind of reticular fiber staining kit for pathological examination, main inclusion:Potassium permanganate 5ml, grass
Sour 3ml, iron alum 3ml, diamino silver hydroxide 2ml, formaldehyde 1ml, chlorauride 5-ml, sodium thiosulfate 2ml.
A kind of using method of the reticular fiber staining kit for pathological examination is:
A. the section preparing is carried out after dewaxing treatment, rinse 1 minute in distilled water;
B. contaminate 2 minutes in liquor potassic permanganate, distillation washing 2 times;
C. after oxalic acid bleaching, distillation washing 2 times;
D. using iron alum aqueous solution mordant dyeing 5 minutes, distillation washing 3 times;
E. diamino silver hydroxide is contaminated 1 minute, distillation washing 3 times;
F. formalin reductase 12 minute, distilled water 1 time are used;
G. mixed colours 3 minutes with chlorogold solution, distilled water 2 times;
H. 3 minutes are fixed with sodium thiosulfate, distillation washing 1 time;
I. use quality concentration is that 95% alcohol is suddenly washed, and until observing clearly till fiber, carries out weight when needing
Counterstain, carries out after the completion of dyeing being dehydrated, transparent and sealing.
Wherein, described liquor potassic permanganate mass concentration is 0.25%;Described oxalic acid mass concentration is 0.8%;Institute
The iron alum aqueous solution mass concentration stated is 1.5%;Described formalin mass concentration is 8%;Described chlorogold solution
Quality solubility is 0.2%;Described sodium thiosulfate mass concentration is 3%.After dyeing according to described using method, ammargenum
By the protein combination in tissue absorption and tissue, it is deposited in tissue and surface through the argent that formaldehyde is reduced into black, uses chlorine
After changing gold toning, then wash away unreduced silver salt with sodium thiosulfate liquid, so that in-house reticular fibre is clearly with black
Color shows.Described dehydration refers to carry out processed using absolute alcohol, and absolute alcohol has very strong water imbibition, and
The hardening of tissue can be made, make the colour developing of fibr tissue more stable.
Embodiment 2:A kind of reticular fiber staining kit for pathological examination, main inclusion:Potassium permanganate 7.5ml,
Oxalic acid 4ml, iron alum 4ml, diamino silver hydroxide 3ml, formaldehyde 2ml, chlorauride 7.5ml, sodium thiosulfate 3.5ml.
A kind of using method of the reticular fiber staining kit for pathological examination is:
A. the section preparing is carried out after dewaxing treatment, rinse 2 minutes in distilled water;
B. contaminate 3 minutes in liquor potassic permanganate, distillation washing 2 times;
C. after oxalic acid bleaching, distillation washing 2 times;
D. using iron alum aqueous solution mordant dyeing 7.5 minutes, distillation washing 4 times;
E. diamino silver hydroxide is contaminated 2 minutes, distillation washing 3 times;
F. reduced 3 minutes with formalin, distilled water 2 times;
G. mixed colours 4 minutes with chlorogold solution, distilled water 2 times;
H. 4 minutes are fixed with sodium thiosulfate, distillation washing 2 times;
I. use quality concentration is that 95% alcohol is suddenly washed, and until observing clearly till fiber, carries out weight when needing
Counterstain, carries out after the completion of dyeing being dehydrated, transparent and sealing.
Wherein, described liquor potassic permanganate mass concentration is 0.3%;Described oxalic acid mass concentration is 0.9%;Described
Iron alum aqueous solution mass concentration be 2.5%;Described formalin mass concentration is 10%;Described chlorogold solution matter
Amount solubility is 0.2%;Described sodium thiosulfate mass concentration is 4%.After dyeing according to described using method, ammargenum quilt
Protein combination in tissue absorption and tissue, is deposited in tissue and surface through the argent that formaldehyde is reduced into black, uses chlorination
After gold toning, then wash away unreduced silver salt with sodium thiosulfate liquid, so that in-house reticular fibre is clearly with black
Show.Described dehydration refers to carry out processed using absolute alcohol, and absolute alcohol has very strong water imbibition, and energy
Make the hardening of tissue, make the colour developing of fibr tissue more stable.
Embodiment 3:A kind of reticular fiber staining kit for pathological examination, main inclusion:Potassium permanganate 10ml, grass
Sour 5ml, iron alum 5ml, diamino silver hydroxide 4ml, formaldehyde 3ml, chlorauride 10ml, sodium thiosulfate 5ml.
A kind of using method of the reticular fiber staining kit for pathological examination is:
A. the section preparing is carried out after dewaxing treatment, rinse 3 minutes in distilled water;
B. contaminate 4 minutes in liquor potassic permanganate, distillation washing 3 times;
C. after oxalic acid bleaching, distillation washing 2 times;
D. using iron alum aqueous solution mordant dyeing 10 minutes, distillation washing 5 times;
E. diamino silver hydroxide is contaminated 3 minutes, distillation washing 3 times;
F. reduced 4 minutes with formalin, distilled water 3 times;
G. mixed colours 5 minutes with chlorogold solution, distilled water 2 times;
H. 5 minutes are fixed with sodium thiosulfate, distillation washing 3 times;
I. use quality concentration is that 95% alcohol is suddenly washed, and until observing clearly till fiber, carries out weight when needing
Counterstain, carries out after the completion of dyeing being dehydrated, transparent and sealing.
Wherein, described liquor potassic permanganate mass concentration is 0.35%;Described oxalic acid mass concentration is 1%;Described
Iron alum aqueous solution mass concentration be 3.5%;Described formalin mass concentration is 12%;Described chlorogold solution matter
Amount solubility is 0.2%;Described sodium thiosulfate mass concentration is 5%.After dyeing according to described using method, ammargenum quilt
Protein combination in tissue absorption and tissue, is deposited in tissue and surface through the argent that formaldehyde is reduced into black, uses chlorination
After gold toning, then wash away unreduced silver salt with sodium thiosulfate liquid, so that in-house reticular fibre is clearly with black
Show.Described dehydration refers to carry out processed using absolute alcohol, and absolute alcohol has very strong water imbibition, and energy
Make the hardening of tissue, make the colour developing of fibr tissue more stable.
Clinical statisticses are tested:
The clinical tissue collecting lymphosarcoma and reticulosarcoma, and it is respectively prepared pathological section, carry out pathology
Check, wherein lymphosarcoma is cut into slices 80 parts, reticulosarcoma is cut into slices 80 parts, is divided into four groups, normal detection group 1, normal detection
Group 2, normal detection group 3, control group, every group contains lymphosarcoma section and each 20 parts of reticulosarcoma section.
Test method:The reticular fibre that 3 groups of normal detected components are not prepared using embodiment 1, embodiment 2, embodiment 3
Staining kit is carried out after dyeing process in electric Microscopic observation, and the pathological section of control group is used Hematoxylin-eosin decoration method
In electric Microscopic observation after being dyeed, Statistical Comparison is carried out to four groups of observation result.
Criterion:
(1) obvious:Substantially, dyeing is high-visible for feature, has good reference property;
(2) obscure:Feature Fuzzy, has part dyeing visible it is impossible to judge;
(3) invalid:Dye-free sign.
Result of the test shows:The staining kit of the present invention has higher sensitive compared with Hematoxylin-eosin decoration method
Degree and specificity, Color is good, and feature substantially, has good potential applicability in clinical practice.
Finally it should be noted that:Above example only in order to technical scheme to be described, is not intended to limit;Although
With reference to the foregoing embodiments the present invention is described in detail, it will be understood by those within the art that:It still may be used
To modify to the technical scheme described in previous embodiment, or equivalent is carried out to wherein some technical characteristics;And
These modifications or replacement, do not make the essence of appropriate technical solution depart from spirit and the model of embodiment of the present invention technical scheme
Enclose.
Claims (5)
1. a kind of reticular fiber staining kit for pathological examination is it is characterised in that kit mainly includes:Potassium permanganate
5-10ml, oxalic acid 3-5ml, iron alum 3-5ml, diamino silver hydroxide 2-4ml, formaldehyde 1-3ml, chlorauride 5-10ml, thio sulphur
Sour sodium 2-5ml.
2. as claimed in claim 1 a kind of reticular fiber staining kit for pathological examination it is characterised in that described
Kit using method is:
A. the section preparing is carried out after dewaxing treatment, distilled water rinses 1-3 minute;
B. 2-4 minute, distillation washing 2-3 time are contaminated in liquor potassic permanganate;
C. after oxalic acid bleaching, distillation washing 2 times;
D. using iron alum aqueous solution mordant dyeing 5-10 minute, distillation washing 3-5 time;
E. diamino silver hydroxide contaminates 1-3 minute, distillation washing 3 times;
F. use formalin reductase 12-4 minutes, distilled water 1-3 time;
G. with chlorogold solution toning 3-5 minute, distilled water 2 times;
H. 3-5 minute, distillation washing 1-3 time are fixed with sodium thiosulfate;
I. use quality concentration is that 95% alcohol is suddenly washed, and until observing clearly till fiber, carries out repeating to contaminate when needing
Color, carries out after the completion of dyeing being dehydrated, transparent and sealing.
3. as claimed in claim 2 a kind of reticular fiber staining kit for pathological examination it is characterised in that described
Liquor potassic permanganate mass concentration is 0.25-0.35%;Described oxalic acid mass concentration is 0.8-1%;Described iron alum water
Concentration of polymer solution is 1.5-3.5%;Described formalin mass concentration is 8-12%;Described chlorogold solution quality is molten
Spend for 0.2%;Described sodium thiosulfate mass concentration is 3-5%.
4. as claimed in claim 2 a kind of reticular fiber staining kit for pathological examination it is characterised in that according to institute
After the using method dyeing stated, reticular fibre is in black.
5. as claimed in claim 2 a kind of reticular fiber staining kit for pathological examination it is characterised in that described
Dehydration refers to carry out processed using absolute alcohol.
Priority Applications (1)
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CN112539986A (en) * | 2020-12-01 | 2021-03-23 | 河南赛诺特生物技术有限公司 | Ferrous sulfate reagent and kit for melanin dyeing |
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CN102589955A (en) * | 2012-01-15 | 2012-07-18 | 中国人民解放军第四军医大学 | Staining reagent kit for detecting tubercle bacillus in body fluid cells |
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CN102589955A (en) * | 2012-01-15 | 2012-07-18 | 中国人民解放军第四军医大学 | Staining reagent kit for detecting tubercle bacillus in body fluid cells |
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CN112539986A (en) * | 2020-12-01 | 2021-03-23 | 河南赛诺特生物技术有限公司 | Ferrous sulfate reagent and kit for melanin dyeing |
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