CN106442074A - Reticular fiber staining kit for pathological examination - Google Patents

Reticular fiber staining kit for pathological examination Download PDF

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Publication number
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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reticular
kit
tissue
pathological examination
minute
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李富亮
刘春英
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N2001/302Stain compositions
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00Sampling; Preparing specimens for investigation
    • G01N1/28Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N2001/305Fixative compositions

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  • 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

A kind of reticular fiber staining kit for pathological examination
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.
CN201610740723.4A 2016-08-28 2016-08-28 Reticular fiber staining kit for pathological examination Pending CN106442074A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
尹有群 等: "改良Gordon-Sweet染色在肝疾病诊断中的应用", 《实用医技杂志》 *
李宝林: "Gordon-Sweet氏网状纤维染色的应用和体会", 《青岛医学院学报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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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Application publication date: 20170222